Boswellia oil, its fractions and compositions for enhancing brain function

ABSTRACT

The present disclosure relates to non-acidic extracts or fractions selected from a  Boswellia  low polar gum resin extract fraction (BLPRE), a  Boswellia  volatile oil fraction (BVOIL), and a  Boswellia  oil fraction (BOIL) and their compositions. BLPRE, BOIL, and BVOIL are each derived from the gum resin of a  Boswellia  species. These compositions are useful for improving mental condition, enhancing brain functions such as cognition, memory, learning, communication and brain health, for treating impaired memory, and for preventing, control or treating memory and cognition related disorders/diseases. Additionally, BOIL, BVOIL, and mixtures of BOIL and BVOIL are useful for enhancing the bioavailability of a biological agent.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application claiming priority to U.S.patent application Ser. No. 13/616,283, filed on Sep. 14, 2012. U.S.patent application Ser. No. 13/616,283 is a continuation-in-part ofinternational application PCT/IN2011/000170, filed on Mar. 14, 2011,which claims priority to Indian patent application 687/CHE/2010, filedMar. 15, 2010; and also a continuation-in-part of internationalapplication PCT/IN2011/000364, filed on May 26, 2011, which claimspriority to Indian patent application 688/CHE/2010, filed May 30, 2010.The entire disclosures of all prior applications are hereby incorporatedby reference in their entirety.

FIELD OF THE DISCLOSURE

The current disclosure provides non-acidic Boswellia low polar gum resinextract fraction (BLPRE), Boswellia volatile oil fraction (BVOIL) andBoswellia oil fraction (BOIL) comprising BLPRE and BVOIL individuallyand their composition(s) obtained by combining with at least onebiological agent or Nootropic agent for the prevention, control andtreatment of brain related diseases comprising Attention-deficitHyperactivity Disorder (ADHD) and memory deficits or to enhance brainfunctions such as cognition, memory, learning and communication.Importantly, the above fractions and compositions of the presentdisclosure help in improving brain health, brain function, and memory.

The current disclosure also provides a non-acidic Boswellia low polargum resin extract fraction (BLPRE), a Boswellia volatile oil fraction(BVOIL), or a Boswellia oil fraction (BOIL) comprising BLPRE and BVOILfor increasing the bioavailability of biological agents. According tothe current disclosure, BLPRE and BVOIL may be prepared by fractionationof BOIL.

BACKGROUND OF THE DISCLOSURE

The gum resin of Boswellia serrata(Burseraceae) plant has long been inuse for the treatment of several diseases by the practitioners ofAyurvedic medicines in the Indian system of medicine. The extract ofBoswellia was found to be a potent anti-inflammatory and anti-arthriticagent. The origin of anti-inflammatory actions of Boswellia gum resinand its extracts has been attributed to a group of triterpene acidscalled boswellic acids that were isolated from the gum resin ofBoswellia serrata. Boswellic acids exert anti-inflammatory actions byinhibiting 5-lipoxygenase (5-LOX). 5-LOX is a key enzyme for thebiosynthesis of leukotrienes from arachidonic acid.3-O-Acetyl-11-keto-β-boswellic acid (AKBA) is biologically the mostactive component among its congeners, it being able to inhibit 5-LOXwith an IC₅₀ of 1.5 μM.

Boswellia gum resin and its extracts also demonstrated significanttherapeutic improvements in human clinical trials confirming theanti-inflammatory effects shown in vitro and in vivo.

Worldwide aging of the population has increased the incidents ofcognitive deficits, such as age-associated memory impairment and seniledementias, and this causes great disruptive impact on the life of theaffected individuals. The “cholinergic hypothesis of learning” played apivotal role in the development of drugs for degenerative diseases.

A disturbance of the cortical cholinergic system accompanied by areduction of choline acetylase (reduced acetylcholine synthesis) isinter alia detectable biochemically in case of neurological diseases.Hence, there is a demand for a medicament whose active substance canameliorate this disturbance and highly available at the target organ(brain) and which is well tolerated, particularly in long-term therapy.

Acetylcholinesterase (AChE) is an important enzyme to hydrolyzeacetylcholine, a neurotransmitter mediating the activity ofparasympathetic nerve, into choline and acetate. AChE is formed in theendoplasmic reticulum, and moves and functions in the cell membrane.AChE is distributed around cholinergic nerve, particularly much at themyoneural junction, and is found in the serum, liver and other tissues.

A wide range of evidence shows that acetylcholinesterase (AChE)inhibition can improve cognitive and mental functions through enhancingcortical cholinergic neurotransmission. The acetylcholinesterase (AChE)inhibitors increase the concentration of acetylcholine and help nervecells to communicate better. The longer acetylcholine remains in thebrain, the longer those cells can call up memories. The earliest knownAChE inhibitors are physostigmine and tacrine.

However, clinical studies show that physostigmine has poor oralactivity, brain penetration and pharmacokinetic parameters while tacrinehas hepatotoxic side effects. Studies were thus focused on finding newtypes of acetylcholinesterase inhibitors that would overcome thedisadvantages of these two compounds.

Donepezil and Rivastigmin inaugurate a new class of AChE inhibitors withlonger and more selective action with manageable adverse effects butstill small improvement of cognitive impairment. Galanthamine (Reminyl),an alkaloid isolated from Galanthus nivalis, is another recentlyapproved AChE inhibitor for the treatment of Alzheimer's. It isselective, long acting, and reversible. Galanthamine produces beneficialeffects in patients. Similarly, huperzine A, a novel Lycopodium alkaloiddiscovered from the Chinese medicinal plant Huperzia serratais a potent,reversible and selective inhibitor of AChE with a rapid absorption andpenetration into the brain in animal tests. It exhibits memory-enhancingactivities in animal and also in clinical trials.

Dementia with Lewy bodies (DLB) is a common cause of dementia. Changesin the acetylcholine system have been reported in brains of patientswith DLB, which provides a rational basis for trials ofacetylcholinesterase inhibitors in DLB.

Current treatment of dementia in Parkinson's disease (PD) is based onthe compensation of profound cholinergic deficiency, as in recentstudies with the cholinesterase inhibitors galantamine, donepezil andrivastigmine. It has also been shown that cholinesterase inhibitors canimprove motor function in PD. The beneficial effect of cholinesteraseinhibitors has been studied on patients suffering from Parkinson'sdisease and dementia.

Studies show that Wernicke-Korsakoff syndrome is associated with apersisting severe anterograde amnesia in which memory is not transferredfrom short-to long-term storage. It is believed to be a consequence of athiamine-deficient state often found in alcohol abusers. The memorydeficit has been attributed to a number of brain lesions (corpusmamillare and dorsomedial nucleus of the thalamus), loss of cholinergicforebrain neurons and serotonin-containing neurons. Many studies andcase-reports suggest efficacy of acetylcholinesterase inhibitors in theWernicke-Korsakoff-associated memory deficit. Studies also suggest thatneurons in the nucleus basalis are at risk in thiamine deficientalcoholic.

The U.S. Pat. No. 5,720,975 relates to the use of incense (olibanum),incense extracts, substances contained in incense, their physiologicallyacceptable salts, their derivatives and their physiological salts, pureboswellic acid, of physiologically acceptable salts, of a derivative, ofa salt of the derivative, for production of a medicament for theprevention or treatment of Alzheimer's disease.

US publication US20060177467A1 relates to the use of the hydrogenationproducts of frankincense (olibanum), its hydrogenated ingredients aswell as physiologically acceptable salts and derivatives thereof andhydrogenated frankincense extracts for the production of a medicamentfor the prophylactic and/or therapeutic treatment of cerebral ischemia,cranial/brain trauma and/or Alzheimer's disease.

There is currently no prior art, to the best of the Applicants'knowledge, relating to the use of Boswellia non-acidic oil fractions andtheir compositions for the prevention, control and treatment of Memoryand Cognition related diseases and enhancing brain functions.

Additionally, there are numerous pharmaceutical ingredients, herbalingredients and biologically active molecules that are effective invitro against a disease condition or disorder. However, several of theseingredients are not effective or not bioavailable in vivo, i.e., afteradministration to warm blooded animals. It is thus important to exploreand identify safe and effective agents that help to increase thebioavailability of such ingredients. As set forth in the presentdisclosure, Boswellia non-acidic oil fractions have been found toincrease bioavailability of a number of extracts, fractions,phytochemicals and compounds originating from plant, animal ormicroorganism sources.

There is currently no prior art, to the best of the Applicants'knowledge, relating to the use of Boswellia non-acidic oil fractions andtheir compositions for increasing the bioavailability of biologicalagents in warm blooded animals.

SUMMARY OF THE DISCLOSURE

Various embodiments of the present disclosure provide use ofcompositions comprising Boswellia non-acidic fraction(s) selected fromBoswellia low polar gum resin extract fraction (BLPRE) having a novelphytochemical composition, Boswellia volatile oil fraction (BVOIL), anda Boswellia oil fraction (BOIL) comprising BLPRE and BVOIL fractions,either individually or in combination. These compositions are useful forimproving brain health and brain functions, which include but are notlimited to cognition, memory, intelligence, motivation, attention,concentration, learning power and better communication. Thesecompositions are also useful to alleviate disease conditions related tocognition and memory deficits and the like.

Various embodiments disclosed herein provide use of Boswellia non-acidicfraction(s) selected from Boswellia low polar gum resin extract fraction(BLPRE), Boswellia volatile oil fraction (BVOIL), and a Boswellia oilfraction (BOIL) comprising BLPRE and BVOIL, either individually or ascompositions, to prevent, control and treat brain relateddiseases/disorders which include but not limited to senile dementia,multi-infarct dementia, dyslexia, aphasia, organic brain syndrome,myasthenia gravis, vascular dementia, mild cognitive impairment (MCI),Lewy body dementia, Wernicke-Korsakoff-syndrome, Alzheimer's,Parkinson's disease, Attention-deficit Hyperactivity Disorder (ADHD),hypoxia, anoxia, cerebrovascular insufficiency, epilepsy, myoclonus andhypocholinergic dysfunctions, to slowdown memory deterioration,functional loss and to treat memory impairment disorders,neurodegenerative disorders, and for controlling blood pressure andblood circulation in the brain.

Various embodiments disclosed herein relate to compositions comprisingat least one component selected from Boswellia low polar gum resinextract fraction (BLPRE), Boswellia volatile oil fraction (BVOIL) andnon-acidic Boswellia oil fraction consisting of BLPRE and BVOIL incombination with at least one component selected from biologicalagent(s), Nootropic agent(s).

In another aspect, the disclosure provides compositions comprising atleast one component selected from Boswellia low polar gum resin extractfraction (BLPRE), Boswellia volatile oil fraction (BVOIL) and non-acidicBoswellia oil fraction (BOIL) consisting of BLPRE and BVOIL incombination with at least one component selected from Boswelliaextract(s), fraction(s), extracts/fractions enriched in one or moreboswellic acids, their salts or derivatives thereof.

In another aspect, the disclosure provides compositions comprising atleast one component selected from Boswellia low polar gum resin extractfraction (BLPRE), Boswellia volatile oil fraction (BVOIL) and non-acidicBoswellia oil fraction (BOIL) in combination with one or more agentsselected from natural antioxidants, anti-inflammatory agents and immunemodulators.

In another embodiment, the disclosure provides Boswellia low polar gumresin extract fraction (BLPRE) for increasing the bioavailability ofbiological agents.

In further embodiments, the disclosure provides compositions comprisingat least one component selected from Boswellia oil (BOIL), Boswelliavolatile oil (BVOIL) and Boswellia low polar gum resin extract (BLPRE)obtained from Boswellia gum resin in combination with a biologicalagent, for increasing the bioavailability of biological agents in warmblooded animals in need thereof.

In various embodiments, the disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or morebiological ingredients or functional ingredients.

In certain embodiments, the disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or morepharmaceutical drugs/synthetic drugs.

In various embodiments, the disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or moreBoswellia derived components.

In various embodiments, the disclosure provides non-acidic Boswelliaderived bioenhancing agents for increasing the bioavailability of one ormore acidic Boswellia derived components.

In certain embodiments, the disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or moreCurcuma derived components.

In various embodiments disclosed herein, a non-acidic Boswellia oilfraction selected from the group consisting of an intact Boswellia oil(BOIL) and a Boswellia volatile oil (BVOIL) may be produced by:

-   -   procuring the gum resin of a plant of the genus Boswellia;    -   extracting said gum resin with a non-polar organic solvent to        produce a non-polar solvent extract solution;    -   washing the non-polar solvent extract solution with an alkali        solution to remove acidic compounds from the non-polar solvent        extract solution;    -   washing the non-polar solvent extract solution successively with        water and brine;    -   evaporating non-polar solvent from the non-polar solvent extract        solution to obtain BOIL as an oily residue; and optionally    -   isolating volatile components from BOIL as BVOIL.

BRIEF DESCRIPTION OF FIGURES

FIGS. 1 and 2 show processes for obtaining BLPRE, BOIL, and BVOIL.

FIG. 3 shows structural formulae 1-9 representing prominent compounds ofBoswellia serrata low polar gum resin extract (BsLPRE).

FIG. 4 shows the HPLC chromatogram depicting the phytochemical profileof the Boswellia serrata low polar gum resin extract (BsLPRE).

FIGS. 5A, 5B, and 5C show bar diagrammatic representation of number ofdays required for learning, latency in finding feed and number of wrongentries respectively obtained during learning phase. The bars 1 to 3represents vehicle treated control, BsLPRE (250 mg/kg) and piracetam(150 mg/kg) respectively. Each bar represent mean±SE, n=8, *p<0.05 and**p<0.01.

FIGS. 6A and 6B show bar diagrammatic representation of latency infinding feed and number of wrong entries respectively obtained duringmemory retention phase. The bars 1 to 3 represents vehicle treatedcontrol, BsLPRE (250 mg/kg) and piracetam (150 mg/kg) respectively. Eachbar represent mean±SE, n=8.

FIG. 7 shows a plot of serum concentration of AKBA after oraladministration of the composition LI13108F containing Boswellia serratalow polar gum resin extract (BsLPRE) and Boswellia serrata extractselectively enriched to 30% 3-O-acetyl-11-keto-β-boswellic acid (AKBA)and composition LI13119F containing Boswellia serrata volatile oilfraction (BsVOIL) and Boswellia serrata extract selectively enriched to30% 3-O-acetyl-11-keto-β-boswellic acid (AKBA) to albino rats at dosesequivalent to 30 mg/kg of AKBA.

FIG. 8 represents a plot of serum concentration of Bisdemethylcurcumin(LI01008) after oral administration of composition (LI13124F1)containing Bisdemethylcurcumin and Boswellia serrata low polar gum resinextract (BsLPRE) in 2:1 ratio at concentration of 450 mg/kg orBisdemethylcurcumin (LI01008) alone at 300 mg/kg body weight.

DETAILED DESCRIPTION A. Definitions

-   -   1. ‘Boswellia oil’ or ‘non-acidic Boswellia extract’ or ‘BOIL’        used herein refers to non-acidic Boswellia gum resin extract        containing non-acidic Boswellia low polar gum resin extract        fraction (BLPRE) and Boswellia volatile oil fraction (BVOIL)        obtained from gum resin of any of the Boswellia species. BOIL        encompasses ‘BVOIL’ and ‘BcOIL,’ as defined below.    -   2. ‘Boswellia serrata oil’ or ‘non-acidic Boswellia serrata        extract’ or ‘BVOIL’ used herein refers to non-acidic Boswellia        serrata gum resin extract containing non-acidic Boswellia        serrata low polar gum resin extract fraction (BsLPRE) and        Boswellia serrata volatile oil fraction (BsVOIL) obtained from        gum resin of the Boswellia serrata species.    -   3. ‘Boswellia carterii oil’ or ‘non-acidic Boswellia carterii        extract’ or ‘BcOIL’ used herein refers to non-acidic Boswellia        carterii gum resin extract containing non-acidic Boswellia        carterii low polar gum resin extract fraction (BcLPRE) and        Boswellia carterii volatile oil fraction (BcVOIL) obtained from        gum resin of the Boswellia carterii species.    -   4. ‘Boswellia low polar gum resin extract fraction’ or        ‘Boswellia low polar gum resin extract’ or ‘BLPRE’ used herein        refers to non-acidic Boswellia gum resin extract oil fraction        comprising sesquiterpenes, diterpenes, triterpenes and other        oily phytochemicals obtained after removing the volatile        components from Boswellia oil obtained from gum resin of any of        the Boswellia species by any of the processes described. BLPRE        encompasses ‘BsLPRE’ and ‘BcLPRE,’ as defined below.    -   5. ‘Boswellia serrata low polar gum resin extract fraction’ or        ‘Boswellia serrata low polar gum resin extract’ or ‘BsLPRE’ used        herein refers to non-acidic Boswellia serrata gum resin extract        oil fraction comprising sesquiterpenes, diterpenes, triterpenes        and other oily phytochemicals obtained after removing the        volatile components from Boswellia oil obtained from gum resin        of Boswellia serrata species by any of the processes described.    -   6. ‘Boswellia carterii low polar gum resin extract fraction’ or        ‘Boswellia carterii low polar gum resin extract’ or ‘BcLPRE’        used herein refers to non-acidic Boswellia carterii gum resin        extract oil fraction comprising sesquiterpenes, diterpenes,        triterpenes and other oily phytochemicals obtained after        removing the volatile components from Boswellia carterii oil        obtained from gum resin of Boswellia carterii species by any of        the processes described.    -   7. ‘Boswellia volatile oil fraction’ or ‘Boswellia volatile oil’        or ‘volatile oil’ or ‘volatile fraction’ or ‘BVOIL’ used herein        refers to the volatile fraction/extract comprising monoterpenes,        sesquiterpenes, volatile oils and other oily phytochemicals        obtained from gum resin of any of the Boswellia species by any        of the processes described. BVOIL encompasses ‘BsVOIL’ and        ‘BcVOIL,’ as defined below.    -   8. ‘Boswellia serrata volatile oil fraction’ or ‘Boswellia        serrata volatile oil’ or ‘serrata volatile oil’ or ‘serrata        volatile fraction’ or ‘BsVOIL’ used herein refers to the        volatile fraction/extract comprising monoterpenes,        sesquiterpenes, volatile oils and other oily phytochemicals        obtained from gum resin of the Boswellia serrata species by any        of the processes described.    -   9. ‘Boswellia carterii volatile oil fraction’ or ‘Boswellia        carterii volatile oil’ or ‘carterii volatile oil’ or ‘carterii        volatile fraction’ or ‘BcVOIL’ used herein refers to the        volatile fraction/extract comprising monoterpenes,        sesquiterpenes, diterpenes, volatile oils and other oily        phytochemicals obtained from gum resin of the Boswellia carterii        species by any of the processes described.    -   10. ‘Gum’ or ‘Gum resin’ or ‘resin’ used herein refers to an        exudate of Boswellia plant species.    -   11. ‘Phytochemical’ refers to a pure or semi-pure compound or        compounds isolated from plants.    -   12. Cognition refers to acquisition, processing and retention of        information.    -   13. Cognition enhancer(s) refers to substance(s) that enhances        concentration and memory.    -   14. Nootropic agent(s) refers to smart drugs, memory enhancers        and cognitive enhancers, dietary supplements, nutraceuticals,        functional ingredients and functional foods that are purported        to improve mental functions such as cognition, memory,        intelligence, motivation, attention and concentration.    -   15. Biological agent(s) refer to one or more agents selected        from biologically active ingredient(s), anti-oxidant(s), dietary        supplements, herbal ingredients, nutraceuticals, functional        ingredients, functional foods and nootropic agents and oil(s)        their mixtures obtained from        plant(s)/animal(s)/microorganism(s)/synthesis or semi synthesis.    -   16. ‘Biologically active ingredient(s)’ refers to any        pharmaceutically or dietetically acceptable active        ingredient(s); compound (s), extract (s), fraction (s),        phytochemical(s), synthetic drug(s) or their salts or mixtures        thereof derived from plants, animals or microorganisms or        obtained by chemical synthesis/semi-synthesis.    -   17. ‘Functional ingredient(s)’ refers to any herbal ingredients,        dietary supplements, antioxidants, vitamins, minerals, amino        acids, fatty acids, essential oils, fish oils, enzymes,        glucosamine, Chondroitin and probiotics or their salts or        mixtures thereof derived from plants or animals or        microorganisms or chemical synthesis or semi-synthesis.    -   18. ‘Bioenhancer(s)’ refers to agents that enhance the        availability of biological agent(s) through one or more        mechanism(s) in warm blooded animals comprising increasing the        bioavailability, enhancing the serum concentration, improving        gastrointestinal absorption, improving systemic utilization,        improving cross over through certain biological barriers such as        respiratory lining, urinary lining, blood brain barrier and        skin.    -   19. ‘Bioenhancing composition(s)’ refer to compositions        comprising Boswellia derived oil fraction as a Bioenhancer in        combination with one or more biological agent(s).    -   20. ‘BSE 85%’ used herein refers to Boswellia serrata extract        standardized to 85% Boswellic acids.    -   21. ‘BCE 85%’ used herein refers to Boswellia carterii extract        standardized to 85% Boswellic acids.    -   22. ‘CLE 95%’ refers to Curcuma longa extract standardized to        95% Curcuminoids.    -   23. ‘CAE 20%’ refers to Curcuma aromatica extract standardized        to 20% Curcuminoids.

B. Use of Boswellia Non-Acidic Extracts in Enhancing Memory and BrainFunction

The gum resin of Boswellia has been very widely used since ancienttimes. The gum resin of various species of Boswellia such as Boswelliaserrata, Boswellia carterii or Boswellia papyrifera is a complex mixturecomprising Boswellia oil fraction (BOIL) containing essentialoil/Boswellia volatile oil fraction (BVOIL) and non-acidic Boswellia lowpolar gum resin extract fraction (BLPRE); boswellic acids, sugars andpolysaccharide fraction. The Boswellia serrata/Boswelliacarterii/Boswellia papyrifera extracts widely available in theinternational markets are acidic fractions separated from the gum resinwhich are standardized to contain 65% or 85% total Boswellic acids bytitrimetric method of analysis. During the execution of commercialprocess for regular Boswellia extracts derived from Boswelliaserrata/Boswellia carterii/Boswellia papyrifera (85% total Boswellicacids), the acidic fraction, which contains predominantly triterpeneacids including Boswellic acids is separated from the rest of gum resincomponents. The sugars and other polymeric materials get separated outinto the aqueous phase during the enrichment process for total Boswellicacids. The remaining water immiscible low polar compounds are separatedas Boswellia oil fraction/extract. These low polar compounds are eitherabsent or present at very low concentration in both, commercialBoswellia extracts standardized to boswellic acids and Boswelliaextracts selectively enriched in 3-O-acetyl-11-keto-β-Boswellic acid(AKBA).

The Boswellia non-acidic oil fractions BOIL, BVOIL, and BLPRE may beobtained in a number of ways. One method of obtaining the Boswellianon-acidic oil fractions BOIL, BVOIL, and BLPRE is outlined in FIG. 1.According to this method, Boswellia gum resin 1 is extracted with anon-polar or water-immiscible solvent to produce a solution of anextract of Boswellia gum resin 2 in a non-polar solvent. The non-polarsolvent may be a non-polar organic solvent, such as 1,2-dichloroethane,hexane, dichloromethane, chloroform, ethyl acetate, n-butanol, or methyliso-butyl ketone (MIBK). Alternatively, the non-polar solvent may be anon-polar inorganic solvent. The non-polar extract solution 2 is thenwashed with an aqueous base to extract boswellic acids and other acidiccomponents into an aqueous layer, leaving a non-polar solvent layercontaining non-acidic components of the extract of Boswellia gum resin.The non-polar solvent is then evaporated to produce a Boswellia oil(BOIL) 3, which contains volatile components and non-volatilecomponents. BOIL 3 is subjected to steam distillation to volatilize thevolatile components of BOIL 3. After removal of the volatile componentsfrom BOIL 3, the remaining non-volatile oil, here referred to asBoswellia low-polar gum resin extract (BLPRE) 4, may be recovered.Similarly, after steam distillation, the volatile components from BOIL 3may be condensed or otherwise recovered as a Boswellia volatile oil(BVOIL) 5. Thus, BOIL 3 contains both volatile and non-volatilecompounds. BLPRE 4 contains non-volatile compounds. BVOIL 5 containsonly volatile compounds.

An alternative method of obtaining the Boswellia non-acidic oilfractions BOIL, BVOIL, and BLPRE is outlined in FIG. 2. According tothis method, Boswellia gum resin 1 is extracted with a polar solvent,which may be alcohol or aqueous alcohol. The resulting polar Boswelliaextract 6 is portioned between an aqueous base solution and a non-polarsolvent to produce an aqueous layer and a non-polar solvent layer of asolution of an extract of Boswellia gum resin 7 in a non-polar solvent.The aqueous layer may be discarded. The non-polar solvent may be anon-polar organic solvent, such as 1,2-dichloroethane, hexane,dichloromethane, chloroform, ethyl acetate, n-butanol, or methyliso-butyl ketone (MIBK). Alternatively, the non-polar solvent may be anon-polar inorganic solvent. The non-polar solvent is then evaporatedfrom non-polar extract solution 7, leaving a Boswellia oil (BOIL) 3,which contains volatile components and non-volatile components. BOIL 3is subjected to vacuum to volatilize the volatile components of BOIL 3.After removal of the volatile components from BOIL 3, the remainingnon-volatile oil, here referred to as Boswellia low-polar gum resinextract (BLPRE) 4, may be recovered. Similarly, after steamdistillation, the volatile components from BOIL 3 may be condensed orotherwise recovered as a Boswellia volatile oil (BVOIL) 5.

Process for Obtaining Non-Acidic Boswellia Oil (BOIL) Fraction:

A representative process for obtaining Boswellia oil comprises:

-   -   a) procuring the gum resin of one or more of the plant(s)        selected from but not limited to Boswellia serrata or Boswellia        carterii or Boswellia papyrifera or mixtures thereof,    -   b) extraction of the gum resin with a water immiscible organic        solvent,    -   c) filtering the extract carefully to remove the insoluble resin        material,    -   d) washing the organic solvent extract repeatedly with an        aqueous alkali solution such as aqueous potassium hydroxide,    -   e) washing the organic layer with water and brine,    -   f) evaporating the organic layer under vacuum and high        temperature to obtain the oily residue (BOIL).        Processes for Obtaining Boswellia Volatile Oil (BVOIL) Fraction:

The process for obtaining Boswellia volatile oil (BVOIL) is throughsteam distillation or using high vacuum from Boswellia gum resin.

A representative process for obtaining Boswellia volatile oil comprises:

-   -   a) procuring the gum resin of Boswellia and    -   b) separating the Volatile oil component by either steam        distillation or distillation under high vacuum, low temperature        from the said gum resin to obtain BVOIL.

In an alternative process,

-   -   a) BOIL is prepared according to the process described above,    -   b) BOIL is then subjected to steam distillation or vacuum        distillation to collect Boswellia volatile oil (BVOIL)        Processes for Obtaining Boswellia Low Polar Gum Resin Extract        (BLPRE) Fraction:

A representative procedure for obtaining Boswellia low polar gum resinextract (BLPRE) comprises:

-   -   a) extraction of the gum resin of Boswellia species with a water        immiscible organic solvent and filtering the extract carefully        to remove the insoluble resin material,    -   b) washing the organic solvent extract repeatedly with an        aqueous alkali solution such as aqueous potassium hydroxide,    -   c) washing the organic layer obtained after the alkali wash,        with water and brine,    -   d) evaporating the said organic layer under vacuum and high        temperature to obtain the oily residue,    -   e) removing the volatile compounds from the said oily residue        under high vacuum and very high temperature to obtain BLPRE.

Another representative procedure for obtaining Boswellia low polar gumresin extract (BLPRE) comprises:

-   -   a) preparing the alcohol or hydroalcohol extract of Boswellia        gum resin,    -   b) partitioning the alcohol extract between an aqueous alkali        solution and a water immiscible organic solvent,    -   c) separation of the organic solvent layer, followed by        evaporation of the solvent to obtain oily residue,    -   d) removal of volatile compounds from the said oily residue        under high temperature and high vacuum to obtain BLPRE.

Yet another representative procedure for obtaining Boswellia low polargum resin extract (BLPRE) comprises:

-   -   a) extracting the gum resin of Boswellia species with alcohol or        hydro alcohol,    -   b) evaporating the organic solvent to an optimum level of total        solids and then    -   c) adjusting the pH to the alkaline side, preferably pH 9-12,    -   d) repeatedly extracting the solution with an organic solvent,    -   e) evaporating the organic solvent under vacuum and high        temperature to obtain the oily residue,    -   f) evaporating the volatiles from the said oily residue under        high vacuum and high temperature to obtain BLPRE as a        non-volatile residue.

A representative procedure for obtaining Boswellia serrata volatile oil(BsVOIL) comprises:

-   -   a) procuring the gum resin of Boswellia serrata.    -   b) separating the Volatile oil component by either steam        distillation or distillation under high vacuum, low temperature        from the said gum resin to obtain BsVOIL.

Yet another representative procedure for obtaining Boswellia carteriivolatile oil (BcVOIL) comprises:

-   -   a) procuring the gum resin of Boswellia carterii.    -   b) separating the Volatile oil component by either steam        distillation or distillation under high vacuum, low temperature        from the said gum resin to obtain BcVOIL.

The representative processes for obtaining Boswellia volatile oil(BVOIL) from Boswellia serrata, Boswellia carterii are described above.However, a similar process or processes can be applied to any of the gumresin obtained from Boswellia species for producing Boswellia volatileoil (BVOIL).

A representative procedure for obtaining Boswellia serrata low polar gumresin extract (BsLPRE) comprises:

-   -   a) Procuring the gum resin of Boswellia serrata.    -   b) extraction with an water immiscible organic solvent and the        insoluble gum materials were separated by filtration and        discarded,    -   c) washing the organic solvent extract repeatedly with dilute        aqueous alkali solution to remove the acidic compounds,    -   d) washing the organic layer successively with water and brine,    -   e) evaporating the organic layer under vacuum at 60-70° C. to        obtain an oily residue,    -   f) the volatile components are then removed from the said oily        residue under high vacuum and very high temperature to obtain a        viscous oil, which is referred herein after as Boswellia serrata        low polar gum resin extract (BsLPRE).

Alternatively, the BsLPRE can also be prepared by a process comprising:

-   -   a) preparing the alcohol or hydroalcohol extract of Boswellia        serrata gum resin,    -   b) partitioning the alcohol extract between an aqueous alkali        solution and a water immiscible organic solvent,    -   c) separation of the organic solvent layer, followed by        evaporating the organic layer under vacuum at 60-70° C. to        obtain an oily residue,    -   d) the volatile components are then removed from the said oily        residue under high vacuum and high temperature to obtain a        viscous oil, which is referred herein after as Boswellia serrata        low polar gum resin extract (BsLPRE).

A representative procedure for obtaining Boswellia carterii low polargum resin extract (BcLPRE) comprises:

-   -   a) procuring the gum resin of Boswellia carterii,    -   b) extracting the gum resin with an water immiscible organic        solvent and the insoluble gum materials were separated by        filtration and discarded,    -   c) washing the organic solvent extract repeatedly with dilute        aqueous alkali solution to remove the acidic compounds,    -   d) washing the organic layer successively with water and brine,    -   e) evaporating the organic layer under vacuum at 60-70° C. to        obtain an oily residue.    -   f) the volatile components are then removed from the said oily        residue under high vacuum and high temperature to obtain a        viscous oil, which is referred herein after as Boswellia        carterii low polar gum resin extract (BcLPRE).

Alternatively, the BcLPRE can also be prepared by process comprising:

-   -   a) preparing the alcohol or hydroalcohol extract of Boswellia        carterii gum resin,    -   b) partitioning the alcohol extract between an aqueous alkali        solution and a water immiscible organic solvent,    -   c) separation of the organic solvent layer, followed by        evaporating the organic layer under vacuum at 60-70° C. to        obtain an oily residue,    -   d) the volatile components are then removed from the said oily        residue under high vacuum and high temperature to obtain a        viscous oil, which is referred herein after as Boswellia        carterii low polar gum resin extract (BcLPRE).

The representative processes for obtaining Boswellia low polar gum resinextract (BLPRE) from Boswellia serrata and Boswellia carterii aredescribed above. However, a similar process or processes can be appliedto any of the gum resin obtained from Boswellia species for producingthe low polar gum resin extract.

In the above processes for obtaining BLPRE, BOIL, and/or BVOIL, thewater immiscible organic solvent used for extraction of a Boswellia gumresin or for partitioning an alcohol extract may be, but is not limitedto, 1,2-dichloroethane, hexane, dichloromethane, chloroform, ethylacetate, n-butanol, methyl iso-butyl ketone (MIBK) or a suitablecombination thereof. The alkali solution used for washing the organicsolvent extract, or partitioning the alcohol extract, can be selectedfrom Group-I or Group-II metal hydroxides, which include, but are notlimited to, Sodium hydroxide, Potassium hydroxide, Calcium hydroxide,Magnesium hydroxide and mixtures thereof.

The said intact Boswellia oil (BOIL) or Boswellia volatile oil (BVOIL)or Boswellia low polar gum resin extract (BLPRE) constitute significantcomponents in Boswellia gum resin. However, it has very limitedcommercial utility and it is mostly discarded as a waste material.Potential utilization of these fractions have been long overdue. It wasfound unexpectedly that Boswellia serrata low polar gum resin extract(BcLPRE), a fraction obtained after removing the volatile compounds fromthe Boswellia serrata oil, has several beneficial properties.

In our earlier Indian patent application 2229/CHE/2008 filed 15 Sep.2008 and PCT application # PCT/IN2009/000505 filed 14 Sep. 2009 wedisclosed synergistic compositions comprising AKBA enriched fraction andBoswellia serrata non-acidic extract (BNRE). BNRE composition and methodof identification are also disclosed.

In our recent Indian patent application 394/CHE/2010 filed 15 Feb. 2010we disclosed non Boswellic acid fraction and its synergisticcompositions.

As a part of developing new agents for improving brain/mental functionand alleviating disease conditions related to cognition and memorydeficits, a large number of plant extracts have been screened for theirinhibitory property on Acetylcholinesterase enzyme activity. The assaywas performed in vitro by the method of Ellman et al., with minormodifications, using acetylthiocholine iodide as a substrate (Lee J. H.,et. al. Arch Pharm Res 2004, 27(1): 53-56). It was found veryunexpectedly that the non-acidic extract, Boswellia serrata oil (BsOIL),Boswellia serrata low polar gum resin extract (BsLPRE) fraction andBoswellia serrata volatile oil (BsVOIL) fractions, were potentinhibitors of acetylcholinesterase in vitro. BsLPRE for example potentlyinhibited acetylcholinesterase enzyme activity in vitro as shown inTable 2. BLPRE's in vitro efficacy against acetylcholinesterase enzymeis comparable to commercial drug Neostigmin. BsLPRE exhibited an IC50value of 37.01 ng/mL compared to 43.29 ng/mL shown by neostigmin. Itsacetylcholinesterase inhibitory activity was also evaluated by a cellbased in vitro assay in Rat pheochromocytoma PC 12 cells. The inhibitoryproperty of BsLPRE on the enzyme activity was assessed in β-amyloidpeptide induced-rat pheochromocytoma PC 12 cells. Rat pheochromocytomaPC 12 cells were equally distributed with phenol red free Dulbecco'smodified Eagle's red medium (DMEM) (Sigma Life Science, USA) containing10% fetal bovine serum (FBS) in 24-well plate. Cells were pretreatedseparately with BLPRE and positive control Neostigmin for 1 h.Thereafter, cells were induced with 1 μg/mL of β-amyloid peptide(Calbiochem, USA) for 24 h at 37° C. After 24 h, cells were collectedand washed twice with 1×PBS by centrifugation at 1200 rpm for 5 min at4° C. The cell extracts were prepared in solubilization buffer and thecell lysates were analyzed for acetylcholine esterase (AChE) activity.The BsLPRE showed 25.3% inhibition at 100 ng/mL concentration, where asNeostigmin showed 49.1% inhibition at 20 ng/mL as summarized in Table 4.

In order to understand the chemical composition of BsLPRE, separation ofBsLPRE was carried out using column chromatography and high performanceliquid chromatography (HPLC), and several diterpenoid and triterpenoidcompounds were isolated. The structures of the compounds were rigorouslycharacterized using ¹H NMR, ¹³C NMR, DEPT, HSQC and HMBC, Mass spectraldata. The compounds so obtained and identified are guiol (1), nephthenol(2), serratol (3), diterpene X (4), lupeol (5), olean-12-ene-3β-ol (6),olean-12-ene-3α-ol (7), lanosta-8,24-diene-3α-ol (8) andurs-12-ene-3α-ol (9) as depicted in FIG. 3. The fraction, Boswelliaserrata low polar gum resin extract (BsLPRE) was then standardized tothree or more of the phytochemical marker compounds selected from 1 to9. The typical results obtained are summarized in the Table 1 and atypical chromatogram depicting the profile of BsLPRE is presented inFIG. 4. However, compositions of BsLPRE or any other Boswellia low polargum resin extract composition (BLPRE) obtained from any other speciesmay vary based on several factors such as Boswellia species used, age ofthe plant, season of collection of gum resin, geographic location andmanufacturing process employed.

The foregoing results manifest that BsLPRE is a novel compositioncomprising unique combination of sesquiterpenoids, diterpenoids andtriterpenoids and other phytochemical(s). A compound tentativelyidentified as diterpene X (4) and compounds guiol (1), nepthenol (2) andLanosta-8, 24-diene-3α-ol (8) are not known to be metabolites ofBoswellia serrata gum resin.

The low polar gum resin extract of these as well as other Boswelliaspecies comprise a composition having some similarity to that ofBoswellia serrata. However, the low polar gum resin extract of Boswelliacarterii (BcLPRE) has shown biological activity and synergistic effectvery similar to that exhibited by BsLPRE as summarized in the followingin vitro and in vivo studies. The experimental studies are discussed inthe examples.

The acetylcholinesterase inhibitory of different boswellic acids wasalso evaluated in both enzyme based assay and cell based assay and theinhibitory activities are summarized in Tables 3 and 5.

Oxidative stress induced increased ROS is critical for neuronal damage,which is a serious complication with regard to brain health.Interestingly, the low polar gum resin extract of Boswelliaserrata(BsLPRE) showed potent inhibition of reactive oxygen species(ROS) generation in RAW 264.7 mouse macrophages (Table 6). In addition,BsLPRE also showed protection from oxidative stress induced cytotoxicdamage of human neuroblastoma cells. In a cell based assay, oxidativestress induced by H₂O, showed potent cytotoxic effect on theproliferation of IMR32 human neuroblastoma cells. However, the treatmentwith BsLPRE significantly attenuates the proliferation index of IMR32human neuroblastoma cells back to the normal level (Table 7). Hence theobservations confirm that the low polar gum resin extract (BLPRE) offersprotection from neuronal damage and support improving brain health.

The in vivo efficacy of BsLPRE on learning and memory improvement wasproven in rats using elevated radial arm maze (RAM) method. Oraladministration of BsLPRE (250 mg/kg) significantly (P<0.01) decreasedthe number of days required to make the rats learned as per set criteriaand significantly (P<0.05) decreased the time taken to find the food bythe learned rats in the elevated RAM model. The positive controlPiracetin (150 mg/kg) also showed significant improvement in spatiallearning like reduction in latency and Number of wrong entries, whencompared with the control group and the results are as stated below(FIGS. 5A to 5C). The test product BsLPRE also significantly improvescognition and memory retention (FIGS. 6A and 6B). These results confirmthe efficacy shown by BsLPRE in vitro and suggest that the use of BsLPREimproves spatial learning and memory retention. According to thesefindings, BsLPRE is a promising candidate for facilitation of learningand memory.

Synergistic Compositions Comprising Boswellia Extracts:

Cell based and enzyme based in vitro anti-acetylcholinesterase studieswere conducted on a broad array of Boswellia extracts standardized toboswellic acids and Boswellia serrata low polar gum resin extract(BsLPRE), in addition to other herbal extracts. The individual extractsand different combination of these extracts were tested for theirefficacy to inhibit acetylcholinesterase enzyme. It was foundsurprisingly that a composition (composition-1) comprising a combinationof 1) a Boswellia serrata extract containing 85% total boswellic acids(BSE85%) and 2) a Boswellia serrata low polar gum resin extract (BsLPRE)showed potent inhibition of acetylcholinesterase (AChE).

Hence, the foregoing shows that BOIL, BVOIL and BLPRE alone as well asin combination with Boswellic acid(s)/Boswellia extract(s) orfractions(s) containing boswellic acid(s)/extracts standardized toboswellic acids/one or more Nootropic agents are potent inhibitors ofacetylcholinesterase and as such can be used for the prevention, controland treatment of cognitive disorders and improving memory andalleviating disease conditions related to cognition and memory deficits.

Pure boswellic acids and commercially available Boswellia serrataextract standardized to 85% boswellic acids have been used todemonstrate the subject matter disclosed herein. However, any Boswelliaserrata standardized to 40%-100% total boswellic acids by titrimetricmethod of analysis or standardized to 30%-100% total boswellic acids byHPLC method of analysis can also be used.

Similarly, a composition (composition-34) containing low polar gum resinextract (BLPRE) in combination with α-mangostin offers better protectionfrom neuronal damage (Table 6) and hence can improve brain health. Inaddition, composition-34 also showed better protection from oxidativestress induced cytotoxic damage of human neuroblastoma cells in a cellbased assay (Table 7). This result further confirms the potential roleof the composition containing BsLPRE in the improvement of brain health.

Different Embodiments Disclosed Herein are as Outlined Below:

In the primary aspect, the disclosure provides non-acidic Boswellia lowpolar gum resin extract (BLPRE) fraction, Boswellia volatile oil (BVOIL)fraction and Boswellia oil fraction (BOIL) comprising BLPRE and BVOILfor improving mental condition/brain health, treating impaired memoryand alleviating memory and cognition related disorders and otherassociated diseases in warm blooded animals.

In the other primary aspect the disclosure provides compositionscomprising at least one fraction selected from Boswellia oil (BOIL),Boswellia volatile oil (BVOIL) and Boswellia low polar gum resin extract(BLPRE) in combination with one or more biological agents or Nootropicagents for improving mental condition/brain health, treating impairedmemory and alleviating memory and cognition related disorders and otherassociated diseases in warm blooded animal.

In another embodiment the disclosure provides, composition comprising atleast one Boswellia derived non-acidic extract/fraction selected fromBoswellia low polar gum resin extract fraction (BLPRE), Boswelliavolatile oil fraction (BVOIL) and non-acidic Boswellia oil fraction(BOIL) in combination with at least one component selected frombiological agents, phytochemicals, vitamins, amino acids, minerals;pharmaceutically or dietetically acceptable excipients, vehicles,carriers and diluents or mixtures thereof for improving mentalcondition/brain health; enhancing brain functions such as cognition,memory, learning, communication; for treating impaired memory, and forpreventing, control or treating memory and cognition relateddisorders/diseases.

In another embodiment the disclosure provides methods for improvingbrain health and brain functions such as cognition, memory, learning,communication or treating impaired memory in a subject or warm bloodedanimal in need thereof, wherein the method comprises supplementing thesaid subject or warm blooded animal with an effective dose of Boswelliaderived non-acidic extract/fraction or their composition(s).

In another embodiment the disclosure provides methods for preventing,control or treating memory and cognition related disorders/diseases in asubject or warm blooded animal in need thereof, wherein the methodcomprises supplementing the said subject or warm blooded animal with aneffective dose of Boswellia derived non-acidic extract/fraction or theircomposition(s).

In another embodiment the disclosure provides methods of preventing,control or treating memory and cognition related disorders/diseases,wherein memory and cognition related disorders/diseases include but notlimited to senile dementia, multi-infarct dementia, dyslexia, aphasia,organic brain syndrome, myasthenia gravis, vascular dementia, mildcognitive impairment (MCI), Attention-deficient Hyperactivity Disorder(ADHD), Lewy body dementia, Wernicke-Korsakoff-syndrome, Alzheimer's,Parkinson's disease, hypoxia, anoxia, cerebrovascular insufficiency,epilepsy, myoclonus and hypocholinergic dysfunctions, memory impairmentdisorders and neurodegenerative disorders.

In another aspect, the disclosure provides Boswellia low polar gum resinextract (BLPRE) fraction, Boswellia volatile oil (BVOIL) fraction andBoswellia oil (BOIL) fraction comprising BLPRE and BVOIL individually ortheir composition(s) comprising useful for the prevention, control andtreatment of brain related diseases comprising Attention-deficitHyperactivity Disorder (ADHD) and memory deficits or to enhance brainfunctions such as cognition, memory, learning and communication.Importantly, the said fractions and compositions of the presentdisclosure help in making the brain healthy.

In another aspect, the disclosure provides compositions comprising atleast one component selected from Boswellia low polar gum resin extractfraction (BLPRE), Boswellia volatile oil fraction (BVOIL) and non-acidicBoswellia oil fraction (BOIL) in combination with at least onepharmaceutically/dietetically acceptable excipients/diluents, furtheroptionally comprising one or more agents selected from naturalantioxidants, anti-inflammatory agents and immune modulators.

In another embodiment the disclosure provides the composition comprisingat least one Boswellia derived non-acidic extract/fraction incombination with at least one pharmaceutically/dietetically acceptableexcipients/diluents, wherein said pharmaceutically or dieteticallyacceptable excipients, carriers, vehicles and diluents include but notlimited to glucose, fructose, sucrose, maltose, lactose, yellow dextrin,white dextrin, silicon dioxide, microcrystalline cellulose powder,calcium stearate, magnesium stearate, sorbitol, stevioside, corn syrup,citric acid, tartaric acid, malic acid, succinic acid, lactic acid,L-ascorbic acid, dl-alpha-tocopherol, glycerin, propylene glycol,glycerin fatty ester, poly glycerin fatty ester, sucrose fatty ester,sorbitan fatty ester, propylene glycol fatty ester, acacia, carrageenan,casein, gelatin, pectin, agar, nicotinamide, calcium pantothenate,calcium salts, pigments, flavors, preservatives, distilled water,saline, aqueous glucose solution, alcohol, propylene glycol andpolyethylene glycol, various animal and vegetable oils, white softparaffin, paraffin and wax.

In another aspect, the disclosure provides Boswellia non acidicextracts/fractions selected from Boswellia low polar gum resin extractfraction (BLPRE), Boswellia volatile oil fraction (BVOIL) and non-acidicBoswellia oil fraction (BOIL) and their compositions to prevent, controland treat brain related diseases/disorders which include but not limitedto senile dementia, multi-infarct dementia, dyslexia, aphasia, organicbrain syndrome, myasthenia gravis, vascular dementia, mild cognitiveimpairment (MCI), Lewy body dementia, Wernicke-Korsakoff-syndrome,Alzheimer's, Parkinson's disease, Attention-deficit HyperactivityDisorder (ADHD), hypoxia, anoxia, cerebrovascular insufficiency,epilepsy, myoclonus and hypocholinergic dysfunctions, memory impairmentdisorders and neurodegenerative disorders

In another aspect, the disclosure provides Boswellia derived non-acidicextract/fraction or their composition(s) for improving mentalcondition/brain health by slowing down memory deterioration, functionalloss, by inhibiting beta-amyloid plaque deposition, by controlling bloodpressure and blood circulation in the brain.

In another aspect, the Nootropic agent(s) used for making thecomposition comprise one or more agent(s) selected from smart drugs,memory enhancers and cognitive enhancers; dietary supplements, herbalingredients, nutraceuticals, functional ingredients and functional foodsthat improve mental functions such as cognition, memory, intelligence,motivation, attention and concentration.

In another aspect, the Nootropic agents can be selected from one or morecomponents selected from the extract(s)/fraction(s)/phytochemicalsderived from herbs including but not limited to Bacopa species, Curcumaspecies or Rosmarinus species.

In another aspect, the herbal ingredients that can be used for preparingcompositions are selected from including but not limited to Boswelliaserrata, Boswellia carterii, Bacopa monniera, Curcuma longa, Withaniasomnifera, Rosmarinus officinalis, Garcinia mangostana, α-mangostin,Annona squamosa and Sphaeranthus indices.

In another aspect of the disclosure, the Nootropic agents can beselected from extract(s)/fraction(s)/phytochemicals, extracts/fractionsenriched in one or more phytochemicals selected from including but notlimited to Bacopa monnieri, Withania somnifera, Emblica officinalis,Centella asiatica; extract or fraction enriched in one or morephytochemicals selected from including but not limited to Bacoside A3,Bacopaside II, Jujubogenin isomer of bacopasaponin C, Bacopasaponin C,Bacopaside I, Bacosine, Apigenin, Luteolin and Sitosterol-D-glucoside,curcumin, demethoxycurcumin, bisdemethoxycurcumin, monodemethylcurcumin,bisdemethylcurcumin, tetrahydrocurcumin, tetrahydrodemethoxycurcumin,tetrahydrobisdemethoxycurcumin and ar-turmerone, carnosic acid,rosmarinic acid, camphor, caffeic acid, ursolic acid, betulinic acid,rosmaridiphenol, rosmanol and their salts thereof.

In yet another aspect, the disclosure provides compositions comprisingtherapeutically effective combination of Boswellia oil (BOIL/Boswelliavolatile oil BVOIL)/Boswellia low polar gum resin extract (BLPRE) incombination with at least one Boswellia derived component selected fromthe extract(s), fraction(s) enriched with one or more boswellicacids/pure boswellic acid compounds or Nootropic agents for improvingmemory, impaired memory and alleviating memory and cognition relateddisorders and other associated diseases.

In another aspect, the disclosure provides compositions for thecognition enhancement achieved through one or more biological actionscomprising inhibition of Acetylcholinesterase, increase inButyrylcholinesterase and inhibition of β-amyloid aggregation.

In yet another aspect, the disclosure further provides compositionscomprising Boswellia oil (BOIL)/Boswellia volatile oil (BVOIL)/Boswellialow polar gum resin extract (BLPRE) and at least one component, derivedfrom gum resin of Boswellia species, which include but not limited toα-boswellic acid, β-boswellic acid, 3-O-acetyl-α-boswellic acid,3-O-acetyl-β-boswellic acid, 3-O-acetyl-11-keto-α-boswellic acid and3-O-acetyl-11-keto-β-boswellic acid or mixtures thereof for improvingmemory, impaired memory and alleviating memory and cognition relateddisorders and other associated diseases.

In yet another important aspect, the disclosure further providescompositions comprising Boswellia serrata low polar gum resin extract(BsLPRE) or BsVOIL or BsOIL and at least one Boswellia derived componentselected from the extracts or fractions enriched in or standardized toone or more compounds derived from the gum resin of Boswellia whichinclude but not limited to α-boswellic acid, β-boswellic acid,3-acetyl-α-boswellic acid, 3-acetyl-β-boswellic acid,3-acetyl-11-keto-α-boswellic acid and 3-acetyl-11-keto-β-boswellic acidor mixtures thereof for improving memory, improving impaired memory andalleviating memory and cognition related disorders and other associateddiseases.

In another aspect, the disclosure further provides compositionscomprising Boswellia serrata low polar gum resin extract (BsLPRE) orBsVOIL or BsOIL and a Boswellia serrata extract standardized to 30-100%total boswellic acids by titrimetric method of analysis or 20-100% totalboswellic acids by HPLC method of analysis.

In preferred aspect, the disclosure further provides compositionscomprising Boswellia serrata low polar gum resin extract (BsLPRE) orBsVOIL or BsOIL and Boswellia serrata extract standardized to 85% totalboswellic acids by titrimetric method of analysis or 65% total boswellicacids by titrimetric method of analysis.

In another preferred aspect, the disclosure provides compositionscomprising Boswellia serrata low polar gum resin extract (BsLPRE) orBsVOIL or BsOIL and Boswellia serrata extract selectively enriched inAKBA concentration varying from 3-99% by HPLC method of analysis.

In other preferred embodiment, the disclosure further provides a processfor producing the Boswellia low polar gum resin extract (BLPRE), whichinclude extraction of the gum resin of Boswellia species with a waterimmiscible organic solvent followed by washing the organic solventextract with an aqueous alkali solution such as aqueous potassiumhydroxide, followed by water and brine, and then finally evaporating theorganic layer under vacuum to obtain an oil, followed by removing thevolatile compounds under high vacuum and temperature to obtain BLPRE.The water immiscible organic solvent can be selected from hexane,chloroform, dichloromethane, ethyl acetate, methyl isobutyl ketone orany other water immiscible solvent or mixtures thereof.

The process for producing the Boswellia serrata low polar gum resinextract (BsLPRE) is variable and the alternative process for examplecomprise, extracting the gum resin with alcohol or hydroalcohol, andthen evaporating the organic solvent to optimum concentration of totalsolids and then adjusting the solution to pH to 9-11, followed byrepeatedly extracting the solution with a low polar organic solvent andthen evaporating the organic solvent followed by removing the volatilesunder vacuum at high temperature to obtain BsLPRE.

In a further embodiment, the Boswellia serrata intact oil can also beused in place of BsLPRE for improving memory, impaired memory andalleviating memory and cognition related disorders and for making thecompositions of the present disclosure.

The water immiscible organic solvent in the above process can beselected from the solvents but not limited hexane, chloroform,dichloromethane, ethyl acetate, methylisobutylketone, tert-butanol orany other water immiscible solvent.

The Boswellia serrate extract standardized to 30-100% total boswellicacids by a titrimetric method of analysis or 20-100% total boswellicacids by HPLC method of analysis can be prepared from the gum resinusing a known procedure or obtained from a group of commerciallyavailable Boswellia serrata extracts standardized to boswellic acids.

In another aspect of the disclosure, the non acidic extracts Boswelliaoil (BOIL), Boswellia volatile oil (BVOIL), Boswellia low polar gumresin extract (BLPRE) used for the demonstration of the disclosure canbe obtained from the Boswellia species selected from Boswellia serrata,Boswellia carterii, Boswellia papyrifera, Boswellia ameero, Boswelliabullata, Boswellia dakielii, Boswellia dioscorides, Boswellia elongata,Boswellia frereana, Boswellia nana, Boswellia neglecta, Boswelliaogadensis, Boswellia pirottae, Boswellia poporiana, Boswellia rime,Boswellia sacra and Boswellia socotrana.

In another aspect of the disclosure one or more of the Curcuma speciesthat can be used for making the compositions of the present disclosurecan be selected from Curcuma longa, Curcuma aromatica, Curcumadomestica, Curcuma aeruginosa, Curcuma albicoma, Curcuma albiflora,Curcuma alismatifolia, Curcuma angustifolia, Curcuma elata, Curcumaferruginea, Curcuma flariflora, Curcuma yunnanensis and Curcuma edoaria.

In yet another aspect of the present disclosure, BOIL, BVOIL or BLPREalone or in combination with one or more Boswellia derived extractsselectively enriched in boswellic acids/commercially available boswellicextract(s) standardized to 50-100% total boswellic acids/Boswelliaserrata extracts wherein AKBA concentration varies from 3-99% HPLCmethod of analysis and optionally contains one or more ofpharmaceutically/nutraceutically/dietically acceptable excipient(s),diluents, salt(s), additive(s), natural antioxidants or naturalanti-inflammatory agents.

In another aspect, the disclosure provides the usage of BOIL, BVOIL orBLPRE alone or their compositions as it is or in comminuted form and/orin unmodified form as granules or powder or paste or the activeingredients are formulated into a solid, semi-solid or liquid dosageform by adding a conventional biologically or pharmaceuticallyacceptable salt(s) or additive(s) or excipient(s).

In yet another aspect, the disclosure provides use of therapeuticallyeffective amount of BOIL, BVOIL or BLPRE alone or their compositionswith one or more biological agents or Nootropic agents foradministration in a specific dosage form such as orally, topically,transdermally, parenterally or in the form of a kit to a subject orpatient in need thereof. Specific dosage form for formulation of thecompositions of the present disclosure include but not limited to oralagents such as tablets, soft capsule, hard capsule, soft gel capsules,pills, granules, powders, emulsions, suspensions, syrups, pellets, food,beverages, concentrated shots, drops and the like; parenteral agentssuch as injections, intravenous drip and the like; suppositories;transdermal agents such as patches, topical creams and gel; ophthalmicagents and nasal agents.

In another aspect, the present disclosure provides compositionscontaining at least one extract/fraction selected from BOIL, BVOIL orBLPRE in combination with one or more functional ingredient(s)comprising herbal ingredients, dietary supplements, antioxidants,vitamins, minerals, amino acids, fatty acids, essential oils, fish oils,enzymes, glucosamine, Chondroitin and probiotics or their salts ormixtures thereof derived from plants or animals or microorganisms orchemical synthesis or semi-synthesis.

In a further aspect, the present disclosure provides BOIL, BVOIL orBLPRE alone or their compositions further optionally combined witheffective amounts of one or more pharmaceutical/nutraceutical/dieticallyacceptable agents including but not limited to antioxidant(s),adaptogen(s), anti-acetylcholinesterase agent(s), anti-inflammatoryagent(s), anti-diabetic agent(s), antiobese agent(s),antiatherosclerotic agent(s), bio-protectants and/or bio-availabilityenhancer(s) and trace metals.

The examples of the biologically/pharmaceutically acceptable carriersemployed in the present disclosure include, but are not limited to,surfactants, excipients, binders, diluents, disintegrators, lubricants,preservatives, stabilizers, buffers and suspensions.

In alternative aspect of the disclosure, the BOIL, BVOIL or BLPRE aloneor their compositions can be optionally delivered in the form ofcontrolled release dosage forms; and by using techniques includingnanotechnology, microencapsulation, colloidal carrier systems and otherdrug delivery systems. The said formulation can be designed for once adaily administration or multiple administrations per day.

In accordance to the present disclosure, the BOIL, BVOIL or BLPRE aloneor their compositions can also be formulated into or added to existingor new food and beverage form(s) and animal feeds as a healthy food orbeverage or feed.

In accordance to the present disclosure, the BOIL, BVOIL or BLPRE aloneor their compositions can also be formulated into or added to existingor new food and beverage form(s) and animal feeds as a healthy food orbeverage or feed for prevention, control and treatment of brain relateddiseases/disorders.

In yet another embodiment, the composition can comprise 10%-99% by theweight of Boswellia serrata derived component selected from theextract(s) and fraction(s) enriched with one or more boswellic acids,pure boswellic acid compounds and mixtures thereof and 90%-10% by weightof Boswellia serrata low polar gum resin extract BsLPRE or BsVOIL orBsOIL.

C. Use of Boswellia Non-Acidic Extracts as Bio-Enhancing Agents

During the search for bioenhancing agents, it was found that non-acidicBoswellia low polar gum resin extract fraction (BLPRE), Boswelliavolatile oil fraction (BVOIL) or Boswellia oil fraction (BOIL)comprising BLPRE and BVOIL enhance the bioavailability of bioactiveagents. The compositions LI13108F containing Boswellia serrata low polargum resin extract (BsLPRE; LI13115) and Boswellia serrata extractselectively enriched to 30% 3-O_acetyl-11-keto-β-boswellic acid (AKBA)and LI13119F containing Boswellia serrata volatile oil fraction (BsVOIL)and Boswellia serrata extract selectively enriched to 30%3-O_acetyl-11-keto-β-boswellic acid (AKBA) were supplemented to AlbinoWistar rats. The control group of animals was supplemented withBoswellia serrata extract selectively enriched to 30% AKBA. Bloodsamples were collected from all animals prior to oral administration oftest products and at 0.5, 1, 2, 4, 8 and 12 hrs after oraladministration. The comparative oral bioavailability of AKBA from theseBoswellia products was evaluated by measuring the serum AKBAconcentration for each test animal using LC-MS.

Surprisingly, both the compositions LI 13108F and LI 13119F showedbetter oral bioavailability with AUCs 14.08 and 11.23 respectivelycompared to AUC 9.825 shown by individual ingredient Boswellia serrataextract containing 30% AKBA (LI 13115). The bioavailability (in terms ofAUC) of LI 13108F is 43.33% more than LI 13115. The bioavailability ofLI 13119F is 14.33% more than that of LI 13115. The study details aresummarized in example-5 and depicted in FIG. 7.

To exert optimal therapeutic efficacy, an active substance should reachsystemic circulation and site of its action in an effectiveconcentration during the desired period. Improving bioavailability andreducing dosage frequency without losing therapeutic benefit is crucialin achieving therapeutic efficacy and patient compliance in chronictreatment regimes. The compositions disclosed herein achieve thisobjective by enhancing the oral bioavailability of AKBA in compositionscontaining BsLPRE.

The bioavailability enhancing effect of BsLPRE was further confirmed byevaluating the composition LI13124F1 containing BsLPRE and a novelcurcumin compound called bisdemethylcurcumin (LI01008) in comparisonwith LI01008 alone in Alibino Wistar rats. Bisdemethylcurcumin is apotent curcuminoid, far superior to other naturally occurringcurcuminoids with respect to antioxidant and other biological activitiescommonly exhibited curcumins. The composition LI13124F1 showed betterbioavailability of LI01008 in serum samples compared to the animalssupplemented with LI01008 alone. The serum samples of animalssupplemented with LI13124F1 showed 75% better bioavailability comparedto the serum samples of the animals supplemented with LI01008 alone. Theexperimental studies are discussed in example-6 and depicted in FIG. 8.

The foregoing thus suggest that the non-acidic Boswellia low polar gumresin extract fraction (BLPRE), Boswellia volatile oil fraction (BVOIL)or Boswellia oil fraction (BOIL) comprising BLPRE and BVOIL enhance thebioavailability of bioactive agents. These bio-enhancing agents thus canbe useful to improve the efficacy and reduce the dose of bioactiveagents.

In an important aspect, the current disclosure provides bioenhancingagents selected from intact Boswellia oil (BOIL), Boswellia volatile oil(BVOIL) and Boswellia low polar gum resin extract (BLPRE) obtained fromBoswellia gum resin of Boswellia species for increasing thebioavailability of biological agents.

In an important aspect, the current disclosure provides compositionscomprising one or more ingredients selected from intact Boswellia oil(BOIL), Boswellia volatile oil (BVOIL) and Boswellia low polar gum resinextract (BLPRE) obtained from Boswellia gum resin of Boswellia speciesin combination with a biological agent for increasing thebioavailability of biological agent.

In another aspect, the current disclosure provides Boswellia derivedbioenhancing agents for improving the bioavailability and/orbio-efficacy of nutraceuticals or dietary supplements is also relevantto animal health besides being important for humans.

In another aspect the current disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or morebiological ingredient(s) or functional ingredient(s).

In another aspect the current disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or moreBoswellia derived components.

In another aspect the current disclosure provides Boswellia derivedbioenhancing agents for increasing the bioavailability of one or moreCurcuma longa derived components.

In another aspect the current disclosure provides the method of usingBoswellia derived bioenhancing agents for enhancing the bioavailabilityof biological agents.

In another aspect, the current disclosure provides bioenhancing agents,which function through one or more of the mechanisms comprisingincreasing the bioavailability, enhancing the serum concentration,improving gastrointestinal absorption, improving systemic utilizationand improving cross over through certain biological barriers likerespiratory lining, urinary lining, blood brain barrier and skin.

In another aspect, the current disclosure provides bio-enhancing agentsBoswellia oil (BOIL), Boswellia volatile oil (BVOIL) and Boswellia lowpolar gum resin extract (BLPRE) derived from the gum resin of Boswelliawhere in the gum resin can be obtained from one or more of the Boswelliaspecies selected from Boswellia serrata, Boswellia carterii andBoswellia papyrifera.

In another aspect the current disclosure provides compositions forbioenhancing the activity of biological agents in warm blooded animalsin need thereof.

In another aspect the current disclosure provides compositionscomprising Boswellia oil (BOIL), Boswellia volatile oil (BVOIL) andBoswellia low polar gum resin extract (BLPRE) for enhancing thebioavailability of nutraceutical or dietary ingredients in warm bloodedanimals in need thereof.

The nutraceutically/dietetically acceptable agents comprise one or moreingredients selected from phytochemicals, Nootropic agents, anti obeseagents, anti-inflammatory agents, anti cholesterol agents, antiarthritic agents, anti diabetic agents, antimicrobial agents, antifungal agents, anti cancer agents, anti hypertensive agents, analgesicagents, anti platelet aggregation agents, anti atherosclerotic agents,antioxidants, anti thrombotic agents, antibiotic agents, anti malarialagents, anti osteoporotic agents, probiotics agents, anti fungal agents,immune potentiating agents, anti viral agents, anti histamines, musclerelaxants, anti depressants, hypnotic agents and their salts thereof.

In another aspect the current disclosure provides composition(s) forincreasing the bioavailability of one or more biological ingredient(s)selected from biologically active ingredient(s), functionalingredient(s), herbal ingredient(s), dietary supplement(s), nutrient(s),anti-oxidant(s), vitamin(s), mineral(s), amino acid(s), and oil(s) theirmixtures obtained fromplant(s)/animal(s)/microorganism(s)/synthesis/semi-synthesis.

The functional ingredient(s) comprise one or more ingredients selectedfrom nutrients, dietary supplements, nutritional ingredients, herbalingredients, phytochemicals, animal proteins, glucosamine, chondroitin,plant proteins, fruit extracts, animal extracts, algae extracts,probiotics and their salts thereof.

The herbal ingredient(s) comprise one or more ingredients selected fromextracts/fractions/phytochemicals and their salts derived from Withaniasomnifera, Bacopa monniera, Boswellia species, Curcuma species, Centellaasiatica, Sphaeranthus indicus, Annona squamosa, Holopteliaintegrifolia, Piper betel, Dolichos biflorus, Moringa oleifera andMurraya koenigii.

The anti-oxidant(s) comprise one or more ingredients selected fromvitamin A, vitamin C, vitamin E, alpha-carotene, trans-beta-carotene,betacryptoxanthin, lycopene, lutein/zeaxanthin, pine bark bioflavonalscomplex, germanium, selenium and zinc.

The vitamin(s) comprise one or more water soluble vitamins selected fromvitamin B1, vitamin B2, niacinamide, vitamin B6, vitamin B12, folic acidand vitamin C; fat-soluble vitamins selected from vitamin A, vitamin D,vitamin E and vitamin K.

The mineral(s) comprise one or more minerals selected from calcium,iron, zinc, vanadium, selenium, chromium, iodine, potassium, manganese,copper and magnesium.

The amino acid(s) comprise one or more amino acids selected from lysine,isoleucine, leucine, threonine, valine, tryptophan, phenylalanine,methionine, L-selenomethionine and their mixtures thereof.

The oil(s) comprise one or more oils selected from omega-3 fatty acid,flaxseed oil, fish oils, krill oil, essential oils and volatile oils.

The biological activity of Boswellia derived compounds/phytochemicalsthat can be enhanced by bioenhancing agents include extracts offractions standardized to one or more boswellic acids selected fromα-Boswellic acid, β-Boswellic acid, 3-O-acetyl-α-Boswellic acid,3-O-acetyl-β-Boswellic acid, 3-O-acetyl-11-keto-α-Boswellic acid,11-keto-β-Boswellic acid and 3-O-acetyl-11-keto-β-Boswellic acid.

In another aspect, the current disclosure provides bio-enhancing agentsselected from Boswellia oil (BOIL), Boswellia volatile oil (BVOIL) andBoswellia low polar gum resin extract (BLPRE) derived from the gum resinof Boswellia for enhancing the bioavailability of extracts/fractionsparticularly standardized to 3-O-acetyl-11-keto-β-Boswellic acid (AKBA).

In another aspect, the current disclosure provides Boswellia derivedagents and compositions for enhancing the bioavailability of thephytochemicals derived from Boswellia species including but not limitedto boswellic acids selected from α-boswellic acid, β-boswellic acid,3-acetyl-α-boswellic acid, 3-acetyl-β-boswellic acid,3-acetyl-11-keto-α-boswellic acid and 3-acetyl-11-keto-β-boswellic acidor mixtures thereof.

The Boswellia species that can be used for producing the oil (BOIL) orvolatile oil (BVOIL) or low polar gum resin extract (BLPRE) from the gumresin comprise one or more species selected from Boswellia serrata,Boswellia carterii, Boswellia payrifera. Boswellia ameero, Boswelliabullata, Boswellia dakielii, Boswellia dioscorides, Boswellia elongata,Boswellia frereana, Boswellia nana, Boswellia neglecta, Boswelliaogadensis, Boswellia pirottae, Boswellia popoviana, Boswellia rime,Boswellia sacra and Boswellia socotrana.

In another aspect, the current disclosure provides Boswellia oil orBoswellia volatile oil or Boswellia low polar gum resin extract forenhancing the bioavailability of one or more Curcuma derivedextracts/fractions/components/phytochemicals that can be enhanced bybioenhancing agents include extracts of fractions standardized toselected from curcumin, demethoxycurcumin, bisdemethoxycurcumin,monodemethylcurcumin, bisdemethylcurcumin, tetrahydrocurcumin,tetrahydrodemethoxycurcumin, tetrahydro bisdemethoxycurcumin andar-turmerone or mixtures thereof.

In another aspect, the current disclosure provides bio-enhancing agentsBoswellia oil (BOIL), Boswellia volatile oil (BVOIL) and Boswellia lowpolar gum resin extract (BLPRE) derived from the gum resin of Boswelliafor enhancing the bioavailability of extracts/fractions particularlystandardized to curcumin or demethoxycurcumin or bisdemethoxycurcumin ormixtures thereof.

In another aspect, the current disclosure provides Boswellia derivedbioenhancing agents and for enhancing the bioavailability of the one ormore phytochemicals derived from Curcuma species selected from curcumin,demethoxycurcumin, bisdemethoxycurcumin, monodemethylcurcumin,bisdemethylcurcumin, tetrahydrocurcumin, tetrahydrodemethoxycurcumin,tetrahydro bisdemethoxycurcumin and ar-turmerone or mixtures thereof.

The Curcumin derived components that can be bioenhanced are derived fromCurcuma longa, Curcuma aromatica, Curcuma domestica, Curcuma aeruginosa,Curcuma albicoma, Curcuma albiflora, Curcuma alismatifolia, Curcumaangustifolia, Curcumaelata, Curcuma ferruginea, Curcuma flaviflora,Curcuma yunnanensis and Curcuma zedoaria.

D. Examples

The following examples, which include various embodiments, will serve toillustrate the practice of the disclosed subject matter, and it shouldbe understood that the particulars shown are by way of example and forpurpose of illustrative discussion of certain embodiments of theinvention; the following examples do not limit the scope of theinvention.

Example 1

A Process for Preparation of the Non-Acidic Boswellia Extract (BOIL)Comprises:

-   -   g) Procuring the gum resin of one or more of the plant(s)        selected from but not limited to Boswellia serrata or Boswellia        carterii or Boswellia papyrifera or mixtures thereof,    -   h) Extraction of the gum resin with a water immiscible organic        solvent,    -   i) Filtering the extract carefully to remove the insoluble resin        material,    -   j) Washing the organic solvent extract repeatedly with an        aqueous alkali solution such as aqueous potassium hydroxide,    -   k) Washing the organic layer with water and brine,    -   l) Evaporating the organic layer under vacuum and high        temperature to obtain the oily residue (BOIL).

Example 2

A Process for Preparation of the Non-Acidic Boswellia Volatile OilFraction (BVOIL) Comprises:

-   -   a) Procuring the gum resin of one or more of the plant(s)        selected from but not limited to Boswellia serrata or Boswellia        carterii or Boswellia papyrifera or mixtures thereof,    -   b) Separating the Volatile oil component by either steam        distillation or distillation under high vacuum and temperature        from the said gum resin to obtain Boswellia volatile oil        fraction (BVOIL).

Example 3

A Process for Preparation of the Non-Acidic Boswellia Low Polar GumResin Extract Fraction (BLPRE) Comprises:

-   -   a) Procuring the gum resin of one or more of the plant(s)        selected from but not limited to Boswellia serrata or Boswellia        carterii or Boswellia papyrifera or mixtures thereof,    -   b) Extraction of the gum resin with a water immiscible organic        solvent,    -   c) Filtering the extract carefully to remove the insoluble resin        material,    -   d) Washing the organic solvent extract repeatedly with an        aqueous alkali solution such as aqueous potassium hydroxide,    -   e) Washing the organic layer with water and brine,    -   f) Evaporating the organic layer under vacuum and high        temperature to obtain the oily residue (Boswellia oil).    -   g) Taking the said oily residue and removing the volatiles under        high vacuum and high temperature to obtain Boswellia low polar        gum resin extract fraction (BLPRE).

Example 4

Representative Procedure for the Preparation of Boswellia serrata LowPolar Gum Resin Extract Fraction (BsLPRE):

The Boswellia serrata gum resin (100 g) was dispersed in 600 mL ofmethyl isobutyl ketone (MIBK) solvent and stirred at room temperaturefor 60 min. The insoluble gum materials were separated by filtration.The MIBK solution was extracted repeatedly with 2% KOH solution (3×200mL) to remove the acidic compounds. The MIBK layer was then washedsuccessively with water (400 mL) and brine (200 mL). The MIBK layer wasevaporated under reduced pressure at 60-70° C. and the volatilecomponents are removed from the oily residue under high vacuum at75-110° C. to obtain BsLPRE as a viscous oil (12 g).

Alternatively, the gum resin (250 g) collected from Boswellia serratawas extracted with methanol (300 mL×3) and the combined methanol extractwas concentrated. The residue (50 g) was dissolved in ethyl acetate (400mL) and extracted thrice with 1N KOH (3×100 mL). The organic layer waswashed with water (2×200 mL) and brine (200 mL) and evaporated to obtainBoswellia oil. The volatile compounds were evaporated from the oil undervacuum at high temperature (75-110° C.) to obtain 22 g of BsLPRE.

The BsLPRE was subjected to column chromatography over normal silica gelusing solvents of increasing polarity starting from hexane tohexane/ethyl acetate mixtures to ethyl acetate. The identical fractionswere combined based on TLC and combined fractions were subjectedindividually to column chromatography over silica gel using mixtures ofhexane/ethyl acetate or hexane/acetone as eluents to obtain purecompounds. Some of the impure fractions were further subjected topreparative HPLC using a reversed phase C18 silica column to obtain purecompounds. The structures were established by analyzing the ¹H NMR, ¹³CNMR, DEPT, HSQC and HMBC and mass spectral data and then comparing thedata with that of known compounds. Nine of the prominent compounds areidentified as guiol (1), nephthenol (2), serratol (3), diterpene X (4),lupeol (5), olean-12-ene-3β-ol (6), olean-12-ene-3α-ol (7),lanosta-8,24-diene-3α-ol (8) and urs-12-ene-3α-ol (9) as depicted inFIG. 3. The pure compounds were then utilized to standardize theBoswellia serrata low polar gum resin extract (BsLPRE) using HPLCmethod. The novel composition of BsLPRE evaluated based on analyticalHPLC method along with the retention times (R) is summarized in Table 1.The HPLC chromatogram for BsLPRE is depicted in FIG. 4.

TABLE 1 Composition of Boswellia serrata low polar gum resin extractfraction (BsLPRE) S. No Test substance R_(t) in min Percentage 1 Guiol(1) 4.5 0.96 2 Nephthenol (2) 7.087 2.01 3 Serratol (3) 8.027 13.32 4Diterpene X (4) 15.777 0.12 5 Lupeol (5) 26.901 0.06 6Olean-12-ene-3β-ol (6) 31.460 1.29 7 Olean-12-ene-3α-ol (7) 33.718 5.368 Lanosta-8,24-diene-3α-ol (8) 35.371 1.34 9 Urs-12-ene-3α-ol (9) 37.2074.55

Example 5

Boswellia serrata Extract Standardized to 50-100% Total Boswellic Acids(Titrimetric Method):

Boswellia serrata extracts standardized to 85% or 65% total boswellicacids are commercially available. These extracts are standardized usingtitrimetric method of analysis. These extracts can be prepared using aknown procedure. For example, by extracting the gum resin of Boswelliaserrata using a water immiscible solvent and then selectively extractingthe acidic compounds from the organic solvent extract using aqueousalkali solution through phase separation. Finally acidification of thealkali solution to precipitate the boswellic acids followed by vacuumdrying to yield Boswellia serrata extract enriched to 85% boswellicacids (BE85%). Boswellia serrata extracts standardized to a selectedconcentration of total boswellic acids in the range of 40-100% bytitrimetric method of analysis or 30-100% by HPLC method of analysis canbe obtained by purification of the gum resin or the extracts or bydilution of higher grade material.

Example 6

Determination of Acetylcholinesterase Inhibitory Activity of BsOIL,BsLPRE, BcLPRE, BsVOIL and Different Boswellic Acid Compound in an InVitro Enzymatic Assay:

Acetylcholinesterase activity is measured using the substrateacetylthiocholine iodide, which is converted to thiocholine. Thereaction of thiocholine with the chromogenic substrateDithionitrobenzoic acid (DTNB) leads to the formation of a yellow anion,Nitrobenzoic acid, which absorbs strongly at 412 nm. Incubation was donefor 10 min.

The AChE assay was performed by the method of Ellman et al, with minormodifications, using acetylthiocholine iodide as a substrate (Lee J. H.,et. al. Arch Pharm Res 2004, 27(1): 53-56). Ellmans reaction mixturecontains 0.5 mM acetylthiocholine iodide and 1 mM5,5′-dithio-bis-(2-nitrobenzoic acid) in a 50 mM sodium phosphate buffer(pH 8.0). The assay mixture contained 50 μl of 50 mM phosphate buffer atpH—8.0, 30 μl of test substance (BsOIL or BsLPRE or BcLPRE or BsVOIL,different boswellic acids and positive control Neostigmin) at variousconcentrations and 20 μl of (100 mU/mL) enzyme. For blanks, enzyme wasreplaced with phosphate buffer. The reaction mixture was mixedthoroughly, 100 μl of Ellman's reagent was added and incubated at roomtemperature for 10 min. The absorbance was measured at 412 nm usingmicroplate reader. The percentage inhibition of enzyme activity wascalculated by comparing OD's of tests wells with that of control wellsusing the following formula. Calculations: %inhibition=[(control−sample)/control]×100. The results ofAcetylcholinesterase inhibitory activity of BsOIL, BsLPRE, BcLPRE,BsVOIL and boswellic acids are summarized in Table 2 and 3.

TABLE 2 Acetylcholinesterase inhibitory activity of BsOIL, BsLPRE,BcLPRE, BsVOIL Name of the % Inhibition at concentrations of IC50compound 10 ng 25 ng 50 ng ng/mL BsOIL 10.65 17.09 32.69 >50 BsLPRE 31.543.7 57.87 37.01 BcLPRE 15.6 23.18 42.56 >50 BsVOIL 16.5 25.49 33.01 >50BSE-85 — 4.22 6.27 Composition-1 42.7 Neostigmin 25.17 37.19 54.69 43.29

TABLE 3 Acetylcholinesterase inhibitory activity of Pure Boswellic acids% Inhibition at concentrations of IC50 Name of the Product 10 ng 25 ng50 ng 100 ng ng/mL 11-keto-β-boswellic — 11.82 16.36 24.55 >100 acid3-O-acetyl-11-keto- — 9.64 16.07 27.5 >100 β-boswellic acid α-boswellicacid — 13.85 21.92 39.23 >100 β-boswellic acid — 16.75 21.23 31.37 >1003-O-acetyl-α- — 14.86 21.28 33.11 >100 boswellic acid 3-O-acetyl-β- —36.29 40.32 52.02 91.77 boswellic acid Neostigmin 25.17 37.19 54.69 —43.29

Example 7

Acetylcholine Esterase Inhibitory Activity of BsOIL, BsLPRE, BcLPRE,BsVOIL and Boswellic Acids in PC 12 Cells:

The inhibitory property on the enzyme activity was assessed in β-amyloidpeptide induced-rat pheochromocytoma PC 12 cells. Rat pheochromocytomaPC 12 cells were equally distributed with phenol red free Dulbecco'smodified Eagle's red medium (DMEM) (Sigma Life Science, USA) containing10% fetal bovine serum (FBS) in 24-well plate. Cells were pretreatedwith test agents (BsOIL, BsLPRE, BcLPRE, BsVOIL and different boswellicacids and positive control Neostigmin) for 1 h. Thereafter, cells wereinduced with 1 μg/mL of β-amyloid peptide (Calbiochem, USA) for 24 h at37° C. After 24 h, cells were collected and washed twice with 1×PBS bycentrifugation at 1200 rpm for 5 min at 4° C. Cell extracts wereprepared in solubilization buffer (10 mM Tris, pH 7.2; 100 mM NaCl, 50mM MgCl2, 1% Triton X-100). The cell extracts were used as samples formeasuring the acetylcholine esterase (AChE) activity.

Cell extract samples (100 μl) were dispensed into each well of 96-wellmicrotitre plate. Fifty micro liters of DTNB (Dithiobisnitro benzoate)solution was added to each well and incubated for 5 min at roomtemperature. After incubation, 50 μl of acetyl choline iodide solutionwas added to each well and absorbance was read immediately at 405 nm for12 min at 2 min intervals. A standard curve was constructed by usingserial concentrations of acetyl cholinesterase (0-100 mU). Total proteinpresent in 100 μl aliquot of cell extract was calculated by Bradfordmethod and the enzyme activity was normalized and expressed as unitactivity per milligram of protein. Efficacy of test samples wasexpressed in terms of percent inhibition of AChE activity and comparedwith a standard drug, Neostigmin as the positive control.

Results:

Table 4 and Table 5 are summary of the acetyl cholinesterase inhibitoryactivities exhibited by various non acidic extracts from Boswelliaserrata and Boswellia carterii (BsOIL, BsLPRE, BcLPRE and BsVOIL) anddifferent boswellic acids. A standard drug, Neostigmin was used as thepositive control for comparing the AChE inhibitory efficacies of theboswellia products.

TABLE 4 Acetyl cholinesterase inhibitory property of BsOIL, BsLPRE,BcLPRE and BsVOIL % inhibition in AChE Test samples Treatment Conc.activity BsOIL 100 ng/ml 19.92 BsLPRE 100 ng/mL 25.31 BcLPRE 100 ng/mL18.67 BsVOIL 100 ng/mL 16.46 Neostigmin 20 ng/ml 49.09

TABLE 5 Comparative efficacy of different boswellic acids in inhibitingAcetyl cholinesterase activity Treatment % inhibition in Test samplesConc. AChE activity 11-keto- -boswellic acid 100 ng/ml 31.393-O-acetyl-keto- -boswellic acid 100 ng/ml 41.67 α-boswellic acid 100ng/ml 38.42  -boswellic acid 100 ng/ml 46.54 3-O-acetyl-α-boswellic acid100 ng/ml 29.22 3-O-acetyl- -boswellic acid 100 ng/ml 37.34 Neostigmin 20 ng/ml 54.65

Example 8

Preparation of composition-1: Composition-1 was prepared by mixing unitdoses of the following components; four parts of Boswellia serrata lowpolar gum resin extract (BsLPRE) (4 g) and one part of Boswellia serrataextract standardized to 85% total Boswellic acids (BSE 85%) (1 g).

Example 9

Composition-2: Composition-2 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata volatile oil(BsVOIL) (1 g) and four parts of Boswellia serrata extract standardizedto 85% total Boswellic acids (BSE 85%) (4 g).

Example 10

Composition-3: Composition-3 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata non acidic oil(BsOIL) (1 g) and four parts of Boswellia serrata extract standardizedto 85% total Boswellic acids (BSE 85%) (4 g).

Example 11

Composition-4: Composition-4 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g) and four parts of Boswellia serrata extractstandardized to 85% total Boswellic acids (BSE 85%) (4 g).

Example 12

Composition-5: Composition-5 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii volatile oil(BcVOIL) (1 g) and three parts of Boswellia carterii extractstandardized to 85% total Boswellic acids (BCE 85%) (3 g).

Example 13

Composition-6: Composition-6 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BcLPRE) (1 g), four parts of Boswellia serrata extract enrichedwith 20% of 3-O-acetyl-11-keto-β-Boswellic acid (AKBA) (4 g).

Example 14

Composition-7: Composition-7 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g), four parts of Boswellia carterii extractenriched with 20% of 3-O-acetyl-11-keto-β-Boswellic acid (AKBA) (4 g).

Example 15

Composition-8: Composition-8 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BcLPRE) (1 g), four parts of Boswellia serrata extract enrichedwith 40% of 3-O-acetyl-11-keto-β-Boswellic acid (AKBA) (4 g).

Example 16

Composition-9: Composition-9 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g), three parts of Boswellia carterii extractenriched with 40% of 3-O-acetyl-11-keto-β-Boswellic acid (AKBA) (3 g).

Example 17

Composition-10: Composition-10 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BcLPRE) (1 g) and three parts of Bacopa monniera standardizedextract (3 g).

Example 18

Composition-11: Composition-10 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata volatile oilfraction (BsVOIL) (1 g) and three parts of Bacopa monniera standardizedextract (3 g).

Example 19

Composition-12: Composition-12 was prepared by mixing unit doses of thefollowing components; one part of non-acidic Boswellia serrata oilfraction (BsOIL) (1 g) and three parts of Bacopa monniera standardizedextract (3 g).

Example 20

Composition-13: Composition-13 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of Bacopa monniera water extract(4 g).

Example 21

Composition-14: Composition-14 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of Bacopa monniera 90% methanolextract (4 g).

Example 22

Composition-15: Composition-15 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and three parts of Bacopa monniera standardizedextract (3 g).

Example 23

Composition-16: Composition-16 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of Bacopa monniera extractstandardized to 25% bacopasaponins (4 g).

Example 24

Composition-17: Composition-17 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g) and four parts of Bacopa monniera extractstandardized to 25% bacopasaponins (4 g).

Example 25

Composition-18: Composition-18 was prepared by mixing unit doses of thefollowing components; one part of Boswellia papyrifera low polar gumresin extract (BpLPRE) (1 g) and four parts of Bacopa monniera extractstandardized to 25% bacopasaponins (4 g).

Example 26

Composition-19: Composition-19 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of Curcuma longa extractstandardized to 95% total Curcuminoids (CLE 95%) (1 g).

Example 27

Composition-20: Composition-20 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata volatile oilfraction (BsVOIL) (1 g) and four parts of Curcuma longa extractstandardized to 95% total Curcuminoids (CLE 95%) (4 g).

Example 28

Composition-21: Composition-21 was prepared by mixing unit doses of thefollowing components; one part of non-acidic Boswellia serrata oilfraction (BsOIL) (1 g) and four parts of Curcuma longa extractstandardized to 95% total Curcuminoids (CLE 95%) (4 g).

Example 29

Composition-22: Composition-22 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and three parts of curcumin (3 g).

Example 30

Composition-23: Composition-23 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and three parts of bisdemethylcurcumin (BDMC) (3g).

Example 31

Composition-24: Composition-24 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of Withania somnifera methanolextract (4 g).

Example 32

Composition-25: Composition-25 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g) and four parts of standardized Withania somniferaextract (4 g).

Example 33

Composition-26: Composition-26 was prepared by mixing unit doses of thefollowing components; one part of Boswellia low polar gum resin extract(BLPRE) (1 g) and four parts of standardized Rosmarinus officinalisextract (4 g).

Example 34

Composition-27: Composition-27 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and four parts of standardized Rosmarinusofficinalis extract (4 g).

Example 35

Composition-28: Composition-28 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g) and four parts of standardized Rosmarinusofficinalis extract (4 g).

Example 36

Composition-29: Composition-29 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BcLPRE) (1 g) and four parts of Rosmarinus officinalis extractstandardized to 30% Rosmarinic acid (RA 30%) (4 g).

Example 37

Composition-30: Composition-30 was prepared by mixing unit doses of thefollowing components; one part of Boswellia carterii low polar gum resinextract (BcLPRE) (1 g) and three parts of Rosmarinus officinalis extractstandardized to 30% Rosmarinic acid (RA 30%) (3 g).

Example 38

Composition-31: Composition-31 was prepared by mixing unit doses of thefollowing components; one part of Boswellia non acidic oil (BOIL) (1 g)and three parts of Garcinia mangostana methanol extract (3 g).

Example 39

Composition-32: Composition-32 was prepared by mixing unit doses of thefollowing components; one part of non-acidic Boswellia serrata oil(BsOIL) (1 g) and three parts of Garcinia mangostana methanol extract (3g).

Example 40

Composition-33: Composition-33 was prepared by mixing unit doses of thefollowing components; one part of Boswellia serrata low polar gum resinextract (BsLPRE) (1 g) and three parts of Garcinia mangostana methanolextract (3 g).

Example 41

Composition-34: Composition-34 was prepared by mixing unit doses of thefollowing components; two parts of Boswellia serrata low polar gum resinextract (BsLPRE) (2 g) and one part of α-mangostin (1 g).

Example 42

Composition-35: Composition-35 was prepared by mixing unit doses of thefollowing components; four parts of Boswellia serrata low polar gumresin extract (BsLPRE) (4 g) and one part of α-mangostin (1 g).

Example 43

Composition-36: Composition-40 was prepared by mixing unit doses of thefollowing components; one part of Boswellia low polar gum resin extractfraction (BLPRE) (1 g) and three parts of Sphaeranthus indicus ethylacetate extract (3 g).

Example 44

Composition-37: Composition-40 was prepared by mixing unit doses of thefollowing components; one part of non acidic Boswellia oil fraction(BOIL) (1 g) and four parts of Sphaeranthus indicus ethyl acetateextract (4 g).

Example 45

Composition-38: Composition-40 was prepared by mixing unit doses of thefollowing components; one part of Boswellia volatile oil fraction(BVOIL) (1 g) and four parts of Sphaeranthus indicus ethyl acetateextract (4 g).

Example 46

Assay for Measuring Reactive Oxygen Species (ROS)

Formation of ROS was measured using of the fluorescent probe DCFH-DA.The method is based on the incubation of the RAW 264.7 mouse macrophageswith DCFH-DA, which diffuses passively through the cellular membrane.Intracellular esterase activity results in the formation of DCFH, whichemits fluorescence when oxidized to 20, 70-dichlorofluorescein (DCF).Briefly, the cells (final concentration 2×10⁶/ml suspension) wereincubated with DCFH-DA (5 mM) in HEPES-buffered (20 mM) HBSS (CaCl₂ 1.26mM, KCL 5.37 mM, KH₂PO₄ 0.44 mM; MgCl₂

0.49 mM, MgSO4 0.41 mM, NaCl 140 mM, NaHCO₃ 4.17 mM, Na₂HPO₄ 0.34 mM)with glucose (5 mM) at 37° C. for 15 min. Following centrifugation, theextracellular buffer with DCFH-DA was exchanged with fresh buffer andthe suspension was mixed gently. The cells (2×10⁶/ml, 125 ml) weretransferred to 250 ml wells (microtiter plate reader, 96 wells)containing 125 ml buffer with the different concentrations of testsamples (α-mangostin, BsLPRE, Composition-34 and Composition-35) inpresence or absence of 100 mM H₂O₂. Fluorescence was recorded usingexcitation wavelength 485 nm, emission wavelength 530 nm in a Modulusluminescence spectrometer (Turner Biosystems, USA) for 120 min. Resultsare calculated as area under the curve (AUC) and the percentage ofinhibition of intracellular ROS generation was calculated from thecultures treated with H₂O₂. The results are summarized in Table 6.

TABLE 6 Anti-oxidant properties of herbal products and theircombinations Inhibition of Reactive Oxygen Species (ROS) generationSerial no Treatments (IC50) 1 α-mangostin 49.817 ug/ml 2 BsLPRE 53.879ug/ml 3 Composition-34 33.342 ug/ml 4 Composition-35 50.776 ug/ml

Example 47

Cell Proliferation Assay

Effect of BsLPRE or α-mangostin or their combination (Composition-34) oncell growth was tested in oxidative stress induced IMR32 humanneuroblastoma cells SW 982 human synovial cells by using MTT based cellproliferation assay. Briefly, IMR32 human neuroblastoma cells werecultivated in Dulbecco's modified Eagle's red medium (DMEM) (Sigma LifeScience, USA) containing 10% fetal bovine serum (FBS). Equal number ofIMR32 cells was seeded in to each well of 96 well microplate andincubated at 37° C. with 5% CO₂. The cells were treated with 250 M H2O2in presence or absence of different concentrations of BsLPRE orα-mangostin or their combination (Composition-34) for 72 h. Controlwells were supplemented with 0.05% DMSO. After 72 h of treatment, equalvolume of MTT reagent (R&D Systems, USA) was added to each well andincubated for 4 h. Thereafter, 50 μl of solublization buffer (R&DSystems, USA) was added to each well to solubilize the colored formazancrystals produced by the reduction of MTT. After 24 h, the opticaldensity was measured at 550 nm using microplate reader (Bio-Rad, USA).In each assay, the vehicle control and the treatments were done inquadruplicates. The average OD obtained in the vehicle control wells isconsidered as the cell proliferation index of 100. The results aresummarized in Table-7 below.

TABLE 7 Protection from oxidative stress induced cytotoxic damage ofhuman neuroblastoma cells Cell proliferation S. No Treatments Treatmentconc. Index 1 Vehicle Control 100.00 2 H2O2 250 uM 78.71 3 α-mangostin10 ng/ml 80.32 25 ng/ml 86.78 50 ng/ml 87.21 100 ng/ml 91.78 4 BsLPRE 10ng/ml 87.81 25 ng/ml 92.24 50 ng/ml 95.19 100 ng/ml 97.30 5Composition-34 10 ng/ml 92.81 25 ng/ml 95.82 50 ng/ml 99.05 100 ng/ml101.33

Example 48

Inhibition of acetylcholinesterase activity by herbal products and theircombinations in Beta Amyloid protein induced PC12 cells: Theacetylcholinesterase inhibitory activity of α-mangostin, BsLPRE andComposition-34, recorded in Table 8, was measured using the proceduredescribed in example 7.

TABLE 8 Inhibition of acetylcholinesterase activity by herbal products %inhibition in Acetylcholinesterase Serial No. Treatments Treatment conc.(AChE) activity 1 α-mangostin 100 ng/ml 26.83 250 ng/ml 29.48 2 BsLPRE100 ng/ml 30.99 250 ng/ml 35.15 3 Composition-34 100 ng/ml 37.97 250ng/ml 40.57

Example 49

Determination of Spatial Learning and Memory Improvement Efficacy ofBsLPRE in Rats Using Elevated Radial Arm Maze Method:

The animal study protocol was approved by institutional animal ethicscommittee. Sprague Dawley rats were acclimatized for one week andhealthy rats were selected for the study. The selected rats werepre-trained in elevated radial arm maze (RAM) adapted rats wereallocated to various treatment groups each containing eight rats. Aftercompletion of pre-training, the oral treatment was initiated to theanimals and continued daily up to two weeks. During this treatment phasethe rats were placed on the RAM for 10 min each to recognize the foodpellets present in the three different colored arms. During training thespatial learning was estimated by measuring various parameters likenumber of days required to learn the task, latency in finding food andnumber of wrong entries/attempts. After this treatment and training theanimals were given rest without treatment or training for one week(3^(rd) week). On 4^(th) week, the animals were treated with allocateddoses of test products and memory retention test was assessed using thesame animals by measuring latency and number of wrong entries. The datawas analyzed using ANOVA followed by a suitable post-hoc test.

Result (Spatial Learning):

Oral administration of BsLPRE (250 mg/kg) significantly (P<0.01)decreased the number of days required to make the rats learned as perset criteria and significantly (P<0.05) decreased the time taken to findthe food by the learned rats in the elevated RAM model. Piracetam showedsignificant improvement in spatial learning represented by reduction inlatency and Number of wrong entries, when compared with the controlgroup and the results are as stated below (FIG. 5A to 5C). The testproduct BsLPRE also significantly improves cognition and memoryretention (FIGS. 6A and 6B). These results suggest that the use ofBsLPRE improves spatial learning and memory retention. According tothese findings, BsLPRE is a promising candidate for facilitation oflearning and memory.

Example 50

Preparation of Boswellia carterii Low Polar Gum Resin Extract (BcLPRE):

The Boswellia carterii gum resin (100 g) was dispersed in 600 mL ofmethyl iso butyl ketone (MIBK) solvent and stirred at room temperaturefor 60 min. The insoluble gum materials were separated by filtration.The MIBK solution was extracted repeatedly with 2% KOH solution (3×200mL) to remove the acidic compounds. The MIBK layer was then washedsuccessively with water (400 mL) and brine (200 mL). The MIBK layer wasevaporated under reduced pressure at 60-70° C. and the volatilecomponents are then removed from the oily residue under vacuum at 75-85°C. to obtain Boswellia carterii low polar gum resin extract or BcLPRE asa viscous oil (9.5 g).

Alternatively, the gum resin (250 g) collected from Boswellia carteriiwas extracted with methanol (300 mL×3) and the combined methanol extractwas concentrated. The residue (50 g) was dissolved in ethyl acetate (400mL) and extracted thrice with 1N KOH (3×100 mL). The organic layer waswashed with water (2×200 mL) and brine (200 mL) and evaporated to obtainBoswellia oil. The volatile compounds were evaporated from the oil undervacuum at 75-85° C. to obtain 17.75 g of BcLPRE.

Example 51

Comparative Bioavailability of 3-O-acetyl-11-keto-β-boswellic Acid(AKBA) from Different Boswellia Products:

Albino Wistar rats were quarantined and healthy rats were selected forthe study. The selected animals were acclimatized for 7 days prior tothe study initiation in the allocated room. Animals employed for thestudy were randomized into various treatment groups, fasted overnight atfree access to water, body weights were noted and individual doses werecalculated based on the body weights. Blood samples were collected fromall animals prior to oral administration of test products and at 0.5, 1,2, 4, 8 and 12 hrs after oral administration. Collected blood sampleswere allowed to clot for 10 min and centrifuged at 4° C. at 1800 g for10 min. The serum samples were deproteinized with 100 μL TCA (20%) and1.8 mL of HPLC grade methanol, centrifuged at 4° C. at 1800 g for 10 minand supernatants were subjected to LCMS analysis for total AKBA. TheComposition LI 13108F comprising Boswellia serrata extract selectivelyenriched to 30% 3-O-acetyl-11-keto-β-boswellic acid (AKBA) (LI 13115)and Boswellia serrata non-volatile oil (BcLPRE) in the ratio 2:1; andcomposition LI13119F comprising Boswellia serrata extract standardizedto 30% AKBA and Boswellia serrata steam distilled oil (BVOIL) in theratio 2:1 showed better oral bioavailability with Area under the curve(AUC) 14.08 and 11.23 respectively compared to individual Boswelliaserrata extract standardized to 30% AKBA (LI 13115) (AUC: 9.825). Thebioavailability [in terms of [AUC] of LI 13108F is 43.33% more than LI13115. The bioavailability of LI 13119F is 14.33% more than that of LI13115. The serum concentration of AKBA in animals of various treatmentgroups at various time points was summarized in Table 9. The serumconcentration against time was plotted and the details are summarized inFIG. 7.

TABLE 9 Serum concentration of AKBA Mean Serum AKBA concentration μg/mLLI 13115 LI 13108F LI 13119F Time (h) (Mean ± SE) (Mean ± SE) (Mean ±SE) 0 0.000 ± 0.00 0.000 ± 0.00 0.000 ± 0.00 0.5 1.200 ± 0.19 1.533 ±0.06 1.805 ± 0.21 1 1.545 ± 0.28 1.853 ± 0.11 1.865 ± 0.32 2 1.980 ±0.45 2.000 ± 0.16 1.750 ± 0.26 4 0.850 ± 0.32 2.147 ± 0.41 1.100 ± 0.078 0.452 ± 0.22 0.520 ± 0.16 0.645 ± 0.13 12 0.095 ± 0.10 0.199 ± 0.140.200 ± 0.20 AUC 9.825 14.0825 (43.33%) 11.2325 (14.33%)

Example 52

Comparative Bioavailability of LI01008 and its Composition:

LI13124F1 Animals (Wistar Rats) were acclimatized for 7 days prior tostudy initiation. Six animals were divided randomly into 2 groups, eachcomprised of 3 animals. The body weights were noted and doses werecalculated based on initial body weights. Animals were treated orallywith 450 mg dose of a composition (LI13124F1) containingbisdemethylcurcumin (LI01008) and BsLPRE (LI13115) in 2:1 ratio or 300mg/kg LI01008 as suspension in 0.5% CMC. Blood samples were collectedbefore treatment and several time intervals after treatment at 0.5, 1,2, 4, 6, 8 and 12 hours, as plotted in FIG. 8. Collected blood sampleswere processed in a refrigerated centrifuge and serum samples weredeproteinized using HPLC grade methanol, mixed thoroughly andcentrifuged to remove precipitated proteins clear supernatants weresubjected for LI01008 estimation by HPLC. The data is summarized intable 10 below.

As per the data, the bioavailability of LI01008 in the compositionLI13124F1 is 75% better compared to that when LI01008 is administeredalone.

TABLE 10 Serum concentration of bisdemethylcurcumin LI 13124F1 LI 01008suspension in 0.5% suspension in 0.5% S. No Time after Admn. CMC CMC 1 00 0 2 0.5 0.1075 0.07 3 1 0.16 0.1085 4 2 0.105 0.078 5 4 0.065 0.035 66 0.032 0.00535 7 8 0.0085 0 8 12 0 0

It will be appreciated to those of ordinary skill in the art thatchanges could be made to the embodiments described above withoutdeparting from the broad inventive concept thereof. It is understood,therefore, that this invention is not limited to the particularembodiments or examples disclosed, but is intended to covermodifications within the objectives and scope of the present invention.

What is claimed is:
 1. A therapeutic method, comprising administering a composition to an individual in need thereof; said composition comprising an oil extract derived from a Boswellia species, said oil extract being free of acidic compounds; said composition further comprising an optional bioactive agent; said oil extract being present in an amount effective to enhance at least one of: a) cognitive function in said individual; and b) bioavailability of said bioactive agent, said oil extract being prepared by a process comprising: extraction of a Boswellia gum resin with a water-immiscible non-polar solvent to produce a non-polar solvent extract; and washing the non-polar solvent extract with aqueous alkali to remove boswellic acids from the non-polar solvent extract to produce an intact Boswellia oil (BOIL); and subjecting said BOIL to distillation to produce a non-volatile Boswellia oil component and a Boswellia volatile oil (BVOIL) recovered as a distillate from said distillation; wherein said oil extract is selected from the group consisting of the non-volatile Boswellia oil component, BVOIL, BOIL, and a mixture thereof.
 2. The therapeutic method of claim 1, wherein said oil extract is present in an amount effective to enhance cognitive function in said individual; wherein said individual suffers from a cognitive disorder selected from the group consisting of impaired memory, senile dementia, multi-infarct dementia, dyslexia, aphasia, myasthenia gravis, vascular dementia, mild cognitive impairment (MCI), Attention-deficient Hyperactivity Disorder (ADHD), Lewy body dementia, Wernicke-Korsakoff-syndrome, Alzheimer's, Parkinson's disease, hypoxia, anoxia, cerebrovascular insufficiency, epilepsy, myoclonus and hypocholinergic dysfunctions, and neurodegenerative disorders.
 3. The therapeutic method of claim 2, wherein said cognitive disorder is impaired memory.
 4. The therapeutic method of claim 1, wherein said composition comprises said oil extract and at least one component selected from the group consisting of a bioactive agent, a phytochemical, a vitamin, an amino acid, a mineral, a pharmaceutically acceptable excipient, a pharmaceutically acceptable vehicle, a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, and a mixture thereof.
 5. The therapeutic method of claim 1, wherein said Boswellia species is selected from the group consisting of Boswellia serrata, Boswellia carterii, Boswellia papyrifera, Boswellia ameero, Boswellia bullata, Boswellia dalzielii, Boswellia dioscorides, Boswellia elongata, Boswellia frereana, Boswellia nana, Boswellia neglecta, Boswellia ogadensis, Boswellia pirottae, Boswellia popoviana, Boswellia rivae, Boswellia sacra, Boswellia socotrana, and mixtures thereof.
 6. The therapeutic method of claim 1, wherein said composition comprises said bioactive agent, said bioactive agent being selected from the group consisting of dietary supplements, herbal ingredients, nutraceuticals, functional ingredients, functional foods, nootropic agents, and mixtures thereof.
 7. The therapeutic method according to claim 6, wherein said bioactive agent is selected from the group consisting of natural antioxidants, neuroprotecting agents, anti-inflammatory agents, immune modulators, and mixtures thereof.
 8. The therapeutic method according to claim 6, wherein said composition comprises said bioactive agent, said bioactive agent being selected from the group consisting of: at least one Boswellia serrata extract containing at least one boswellic acid, a boswellic acid compound, and a mixture thereof.
 9. The therapeutic method according to claim 1, wherein said composition comprises said bioactive agent, said bioactive agent being selected from the group consisting of: at least one Boswellia serrata extract containing between about 20% and about 100% by weight of total boswellic acids.
 10. The therapeutic method according to claim 1, wherein said composition comprises said bioactive agent, said bioactive agent being selected from the group consisting of: at least one Curcuma longa extract containing at least one curcuminoid, a curcuminoid compound, and a mixture thereof.
 11. The therapeutic method according to claim 1, wherein said composition comprises said bioactive agent, said bioactive agent being selected from the group consisting of said nootropic agents; said nootropic agents being selected from the group consisting of: at least one extract selected from the group consisting of Bacopa monnieri, Withania somnifera, Emblica officinalis, and Centella asiatica; at least one phytochemical selected from the group consisting of Bacoside A3, Bacopaside II, Jujubogenin isomer of bacopasaponin C, Bacopasaponin C, Bacopaside I, Bacosine, Apigenin, Luteolin and Sitosterol-D-glucoside, curcumin, demethoxycurcumin, bisdemethoxycurcumin, monodemethylcurcumin, bisdemethylcurcumin, tetrahydrocurcumin, tetrahydrodemethoxycurcumin, tetrahydrobisdemethoxycurcumin and ar-turmerone, carnosic acid, rosmarinic acid, camphor, caffeic acid, ursolic acid, betulinic acid, rosmaridiphenol, rosmanol, salts thereof, and mixtures thereof.
 12. The therapeutic method according to claim 1, wherein said composition comprises at least one excipient selected from the group consisting of surfactants, binders, disintegrators, lubricants, preservatives, stabilizers, buffers and suspensions.
 13. The therapeutic method according to claim 1, wherein said composition is an oral dosage form.
 14. The therapeutic method according to claim 1, wherein said composition is a controlled release dosage form.
 15. The therapeutic method according to claim 1, wherein said composition contains: an effective amount of said bioactive agent; and said oil extract derived from said Boswellia species in an amount which is effective for enhancing the bioavailability of said bioactive agent.
 16. The therapeutic method according to claim 15, wherein said Boswellia species is selected from the group consisting of Boswellia serrata, Boswellia carterii, Boswellia papyrefera, and mixtures thereof.
 17. The therapeutic method according to claim 15, wherein said composition contains said oil extract in an amount which is effective for enhancing the bioavailability of said bioactive agent in a warm blooded animal in need thereof.
 18. The therapeutic method according to claim 17, wherein said bioactive agent is at least one agent selected from the group consisting of a functional ingredient, an herbal ingredient, a dietary supplement, a nutrient, an antioxidant, a vitamin, a mineral, an amino acid, an oil, and mixtures thereof.
 19. The therapeutic method according to claim 17, wherein the bioactive ingredient comprises one or more ingredients selected from herbal ingredients, dietary supplements, antioxidants, vitamins, minerals, amino acids, fatty acids, essential oils, fish oils, enzymes, glucosamine, chondroitin and probiotics or their salts or mixtures.
 20. The therapeutic method according to claim 17, wherein the bioactive ingredient comprises one or more ingredients selected from the group consisting of extracts or phytochemicals, said extracts or phytochemicals being derived from Withania somnifera, Bacopa monniera, Boswellia species, Curcuma species, Centella asiatica, Sphaeranthus indicus, Garcinia mangostana, Annona squamosa, Holoptelia integrifolia, Piper betel, Dolichos biflorus, Moringa oleifera, Murraya koenigii and mixtures thereof.
 21. The therapeutic method according to claim 18, wherein the herbal ingredient is derived from a Boswellia species, and includes one or more boswellic acids selected from the group consisting of α-boswellic acid, β-boswellic acid, 3-O-acetyl-α-boswellic acid, 3-O-acetyl-β-boswellic acid, 3-O-acetyl-11-keto-β-boswellic acid, and 3-O-acetyl-11-keto-β-boswellic acid.
 22. The therapeutic method according to claim 20, wherein the herbal ingredient is derived from a Boswellia species, and includes an extract comprising 3-acetyl-11-keto-β-boswellic acid.
 23. The therapeutic method according to claim 20, wherein the herbal ingredient is derived from a Curcuma species, and includes a component selected from the group consisting of curcumin, demethoxycurcumin, bisdemethoxycurcumin, monodemethylcurcumin, bisdemethylcurcumin, tetrahydrocurcumin, tetrahydrodemethoxy-curcumin, tetrahydrobisdemethoxycurcumin, ar-turmerone and mixtures thereof.
 24. The therapeutic method according to claim 20, wherein the herbal ingredient is derived from a Curcuma species, and includes curcumin.
 25. The therapeutic method according to claim 18, wherein the antioxidant comprises one or more ingredients selected from the group consisting of vitamin A, vitamin C, vitamin E, alpha-carotene, trans-beta-carotene, betacryptoxanthin, lycopene, lutein, zeaxanthin, pine bark bioflavonals complex, germanium, selenium and zinc.
 26. The therapeutic method according to claim 18, wherein the vitamin comprises: a water-soluble vitamin selected from the group consisting of vitamin B1, vitamin B2, niacinamide, vitamin B6, vitamin B12, folic acid, vitamin C, and mixtures thereof; a fat-soluble vitamin selected from the group consisting of vitamin A, vitamin D, vitamin E and vitamin K, and mixtures thereof; or a mixture of said water-soluble vitamin and said fat-soluble vitamin.
 27. The therapeutic method according to claim 18, wherein the mineral comprises one or more minerals selected from the group consisting of calcium, iron, zinc, vanadium, selenium, chromium, iodine, potassium, manganese, copper, and magnesium.
 28. The therapeutic method according to claim 18, wherein the amino acid comprises one or more amino acids selected from the group consisting of lysine, isoleucine, leucine, threonine, valine, tryptophan, phenylalanine, methionine, and 1-selenomethionine.
 29. The therapeutic method according to claim 18, wherein the oil comprises one or more oils selected from the group consisting of omega-3 fatty acids, flaxseed oil, fish oils, essential oils and volatile oils.
 30. The therapeutic method according to claim 15, wherein said Boswellia species is selected from the group consisting of Boswellia ameero, Boswellia bullata, Boswellia dalzielii, Boswellia dioscorides, Boswellia elongata, Boswellia frereana, Boswellia nana, Boswellia neglecta, Boswellia ogadensis, Boswellia pirottae, Boswellia popoviana, Boswellia rivae, Boswellia sacra and Boswellia socotrana.
 31. A therapeutic method, comprising administering a composition to an individual in need thereof, said composition comprising an effective amount of a bioactive agent and an oil extract derived from a Boswellia species; wherein said composition contains said oil extract in an amount which is effective for enhancing the bioavailability of said bioactive agent; said oil extract being prepared by a process comprising: extraction of a Boswellia gum resin with a water-immiscible non-polar solvent to produce a non-polar solvent extract; washing the non-polar solvent extract with aqueous alkali to remove boswellic acids from the non-polar solvent extract to produce an intact Boswellia oil (BOIL); and subjecting said BOIL to distillation to produce a non-volatile Boswellia oil component and a Boswellia volatile oil (BVOIL) recovered as a distillate from said distillation; wherein said oil extract is selected from the group consisting of the non-volatile Boswellia oil component, BVOIL, BOIL, and a mixture thereof.
 32. The therapeutic method according to claim 31, wherein said bioactive agent is derived from a Boswellia species, and includes one or more boswellic acids selected from the group consisting of α-boswellic acid, β-boswellic acid, 3-O-acetyl-α-boswellic acid, 3-O-acetyl-β-boswellic acid, 3-O-acetyl-11-keto-α-boswellic acid and 3-O-acetyl-11-keto-β-boswellic acid.
 33. The therapeutic method according to claim 31, wherein said bioactive agent is derived from a Curcuma species, and includes a component selected from the group consisting of curcumin, demethoxycurcumin, bisdemethoxycurcumin, monodemethyl-curcumin, bisdemethylcurcumin, tetrahydro curcumin, tetrahydrodemethoxycurcumin, tetrahydrobisdemethoxycurcumin, ar-turmerone and mixtures thereof. 